Chapter 20

Biotechnology
Lecture Outline
Overview: The DNA Toolbox
In 1995, researchers sequenced the entire genome of a free-living organism, the bacterium Haemophilus influenzae. A mere 12 years later, genome sequencing was under way for more than 2, s!ecies. "y 2 #, researchers had com!letely sequenced hundreds of !ro$aryotic genomes and do%ens of eu$aryotic ones, including all & billion base !airs of the human genome. 'a!id advances in ()A technology*methods of wor$ing with and mani!ulating ()A*had their roots in the 19# s. A $ey accom!lishment was the invention of techniques for ma$ing recombinant DNA, ()A molecules formed when segments of ()A from two different sources*often different s!ecies*are combined in vitro. +cientists also have !owerful techniques for analy%ing genes and gene e,!ression. -uman lives are greatly affected by biotechnology, the mani!ulation of organisms or their com!onents to ma$e useful !roducts. o "iotechnology includes such early !ractices as selective breeding of farm animals and the use of microorganisms to ma$e wine and cheese. o .oday, biotechnology also encom!asses genetic engineering, the direct mani!ulation of genes for !ractical !ur!oses. ()A technology is now a!!lied in areas ranging from agriculture to criminal law to medical diagnosis, but many of its most im!ortant achievements are in basic research.

Concept 20.1 DNA cloning yiel m!ltiple copie o" a gene or other DNA egment.
.o study a !articular gene, scientists needed to develo! methods to isolate the small, welldefined !ortion of a chromosome that contains the gene of interest. .echniques for DNA cloning enable scientists to !re!are multi!le identical co!ies of welldefined segments of ()A. /ne common a!!roach to cloning !ieces of ()A uses bacteria, usually Esherichia coli, whose chromosome is a large circular ()A molecule. In addition, bacteria have pla mi# , small circular ()A molecules with a small number of genes that re!licate inde!endently from the chromosome. /ne basic cloning technique begins with the insertion of a 0foreign1 gene into a bacterial !lasmid to !roduce a recombinant ()A molecule.
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.he !lasmid is returned to a bacterial cell, !roducing a recombinant bacterium, which re!roduces to form a clone of genetically identical cells. 2very time the bacterium re!roduces, the recombinant !lasmid is re!licated as well. .he !roduction of multi!le co!ies of a single gene is called gene cloning. 3ene cloning is useful for two basic !ur!oses4 to ma$e many co!ies of a !articular gene and to create a !rotein !roduct. o Isolated co!ies of a cloned gene may enable scientists to determine the gene5s nucleotide sequence or !rovide an organism with a new metabolic ca!ability, such as !est resistance. o Alternatively, a !rotein with medical uses, such as human growth hormone, can be harvested in large quantities from cultures of bacteria carrying the cloned gene for the !rotein. o 6ost !rotein-coding genes e,ist in only one co!y !er genome, so the ability to clone rare ()A fragments is very valuable. 3ene cloning and genetic engineering were made !ossible by the discovery of re triction en$yme that cut ()A molecules at s!ecific locations. In nature, bacteria use restriction en%ymes to cut foreign ()A, to !rotect themselves against !hages or other bacteria. 'estriction en%ymes are very s!ecific, recogni%ing short ()A nucleotide sequences, or re triction ite , and cutting both ()A strands at s!ecific !oints within these sequences. o "acteria !rotect their own ()A by methylating the sequences recogni%ed by these en%ymes. o 2ach restriction en%yme cleaves a s!ecific sequence of bases. o "ecause the target sequence usually occurs 7by chance8 many times on a long ()A molecule, an en%yme ma$es many cuts. o A given restriction en%yme yields the same set of re triction "ragment when it cuts a s!ecific ()A molecule. 'estriction en%ymes cut the covalent sugar-!hos!hate bac$bones of both strands, often in a staggered way that creates single-stranded tic%y en# . .hese e,tensions can form hydrogen-bonded base !airs with com!lementary single-stranded stretches 7stic$y ends8 on other ()A molecules cut with the same restriction en%yme. .hese ()A fusions can be made !ermanent by DNA liga e, which seals the strand by cataly%ing the formation of covalent bonds to close u! the sugar-!hos!hate bac$bone. 'estriction en%ymes and ()A ligase can be used to ma$e a stable recombinant ()A molecule, with ()A that has been s!liced together from two different organisms. 'ecombinant !lasmids are !roduced when restriction fragments from foreign ()A are s!liced into !lasmids. .he original !lasmid used to !roduce recombinant ()A is called a cloning vector, defined as a ()A molecule that can carry foreign ()A into a cell and re!licate there. "acterial !lasmids are widely used as cloning vectors because they can easily be isolated from bacteria, mani!ulated to form recombinant !lasmids by in vitro insertion of foreign ()A, and then reintroduced into bacterial cells.

Restriction enzymes are used to make recombinant DNA.

Eukaryotic genes can be cloned in bacterial plasmids.

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"acterial cells that carry the recombinant !lasmid re!roduce ra!idly, re!licating the inserted foreign ()A. Imagine that researchers are interested in studying the -globin gene in a !articular s!ecies of hummingbird to see whether this o,ygen-carrying !rotein is different from its counter!art in less metabolically active s!ecies. .he first ste! is the isolation of hummingbird genomic ()A, which contains the -globin gene, from hummingbird cells. 'esearchers also isolate the chosen vector, a !articular bacterial !lasmid from E. coli cells. .he !lasmid carries two useful genes, ampR, which confers resistance to the antibiotic am!icillin, and lacZ, which encodes the en%yme 9-galactosidase that cataly%es the hydrolysis of lactose. 9-galactosidase can also hydroly%e a synthetic mimic of lactose called :-gal to form a blue !roduct. .he !lasmid has a single recognition sequence, within the lacZ gene, for the restriction en%yme used. "oth the !lasmid and the hummingbird ()A are digested with the same restriction en%yme. .he fragments are mi,ed together, allowing base !airing between their com!lementary stic$y ends. ()A ligase is added to !ermanently ;oin the base-!aired fragments. +ome of the resulting recombinant !lasmids contain hummingbird ()A fragments< one fragment carries the -globin gene. o .his ste! also generates other !roducts, such as !lasmids containing several hummingbird ()A fragments, a combination of two !lasmids, or a re;oined, nonrecombinant version of the original !lasmid. .he ()A mi,ture is mi,ed with bacteria that have a mutation in the lacZ gene on their own chromosome, ma$ing them unable to hydroly%e lactose or :-gal. .he bacteria ta$e u! foreign ()A by transformation. o +ome cells acquire a recombinant !lasmid carrying a gene, while others may ta$e u! a nonrecombinant !lasmid, a hummingbird ()A fragment, or nothing at all. .he transformed bacteria are !lated on a solid nutrient medium containing am!icillin and :gal. /nly bacteria that have the am!icillin-resistance 7 ampR8 !lasmid grow. 2ach re!roducing bacterium forms a clone by re!eating cell divisions, thus generating a colony of cells on the agar. .he :-gal in the medium is used to identify !lasmids that carry foreign ()A. o "acteria with !lasmids lac$ing foreign ()A stain blue when 9-galactosidase from the intact lacZ gene hydroly%es :-gal. o "acteria with !lasmids containing foreign ()A inserted into the lacZ gene are white because they lac$ 9-galactosidase. In the final ste!, thousands of bacterial colonies with foreign ()A are sorted to find those that contain the gene of interest. In the 0shotgun1 cloning a!!roach described above, a mi,ture of fragments from the entire genome is included in thousands of different recombinant !lasmids.
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Cloned genes are stored in DNA libraries.

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A com!lete set of recombinant !lasmid clones, each carrying co!ies of a !articular segment from the initial genome, forms a genomic library. In addition to !lasmids, certain bacterio!hages are common cloning vectors for ma$ing genomic libraries. o =ragments of foreign ()A can be s!liced into a !hage genome using a restriction en%yme and ()A ligase. o An advantage of using !hage as vectors is that !hage can carry larger ()A inserts than !lasmids can. o .he normal infection !rocess allows the !roduction of many new !hage !articles, each carrying the foreign ()A. o A genomic library made using !hage is stored as a collection of !hage clones. "ecause restriction en%ymes do not recogni%e gene boundaries, some genes in either of these ty!es of genomic library are cut and divided u! among two or more clones. Bacterial arti"icial chromo ome &BAC' are used as vectors for library construction. "A>s are large !lasmids containing only the genes necessary to ensure re!lication and ca!able of carrying inserts of 1 ?& $b. .he very large insert si%e minimi%es the number of clones that are needed to ma$e u! the genomic library, but it ma$es them more difficult to wor$ with. o "A> clones are usually stored in multiwelled !lastic !lates, with one clone !er well. A more limited $ind of gene library can be develo!ed by starting with m')A e,tracted from cells. o .he en%yme reverse transcri!tase is used in vitro to ma$e single-stranded ()A transcri!ts of the m')A molecules. o .he m')A is en%ymatically digested, and a second ()A strand com!lementary to the first is synthesi%ed by ()A !olymerase. o .his double-stranded ()A is called complementary DNA 7cDNA8. o =or creating a library, c()A is modified by the addition of restriction sites at each end and then inserted into vector ()A. o A cDNA library re!resents that !art of a cell5s genome that was transcribed in the starting cell from which the m')A was isolated. If a researcher wants to clone a gene but is unsure in what cell ty!e it is e,!ressed or unable to obtain that cell ty!e, a genomic library will li$ely contain the gene. A researcher interested in the regulatory sequences or introns associated with a gene needs to obtain the gene from a genomic library. o .hese sequences are missing from the !rocessed m')As used in ma$ing a c()A library. A researcher interested in only the coding sequence of a gene can obtain a stri!!ed-down version of the gene from a c()A library. o .his is an advantage if a researcher wants to study the genes res!onsible for the s!eciali%ed functions of a !articular $ind of cell. o "y ma$ing c()A libraries from cells of the same ty!e at different times in the life of an organism, one can trace changes in the !atterns of gene e,!ression. .he researcher screens all the colonies with recombinant !lasmids for a clone of cells containing the hummingbird -globin gene.

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/ne technique, n!cleic aci# hybri#i$ation, de!ends on base !airing between the gene and a com!lementary sequence on a short, single-stranded nucleic acid, a n!cleic aci# probe. Identifying the sequence of the ')A or ()A !robe de!ends on $nowledge of at least !art of the sequence of the gene of interest. A radioactive or fluorescent tag is used to label the !robe, which hydrogen-bonds s!ecifically to com!lementary single strands of the desired gene. .he clones in the hummingbird genomic library have been stored in a multiwell !late. If a few cells from each well are transferred to a defined location on a membrane made of nylon or nitrocellulose, a large number of clones can be screened simultaneously for the !resence of ()A com!lementary to the ()A !robe. /nce the location of a clone carrying the -globin gene has been identified, cells from that colony can be grown in order to isolate large amounts of the -globin gene. o .he cloned gene itself can be used as a !robe to identify similar or identical genes in ()A from other sources, such as other s!ecies of birds. .he !rotein !roduct of a cloned gene can be created in either bacterial or eu$aryotic cells, for research !ur!oses or for !ractical a!!lications. Inducing a cloned eu$aryotic gene to function in bacterial host cells can be difficult because certain as!ects of gene e,!ression are different in eu$aryotes and bacteria. /ne way around this is to insert an expre ion vector, a cloning vector containing a highly active bacterial !romoter, u!stream of the restriction site. .he bacterial host cell recogni%es the !romoter and !roceeds to e,!ress the foreign gene that has been lin$ed to it. o +uch e,!ression vectors allow the synthesis of many eu$aryotic !roteins in bacterial cells. .he !resence of noncoding introns in eu$aryotic genes may !revent the correct e,!ression of these genes in bacteria, which lac$ ')A-s!licing machinery. .his !roblem can be surmounted by using a c()A form of the gene, which includes only the e,ons. 6olecular biologists can avoid incom!atibility !roblems by using eu$aryotic cells as hosts for cloning and e,!ressing eu$aryotic genes. @east cells, single-celled fungi, are as easy to grow as bacteria and, unli$e most eu$aryotes, have !lasmids. +cientists have constructed recombinant !lasmids that combine yeast and bacterial ()A and can re!licate in either ty!e of cell. +cientists have !roduced yea t arti"icial chromo ome 7(AC 8 that combine the essentials of a eu$aryotic chromosome 7an origin site for re!lication, a centromere, and two telomeres8 with foreign ()A. .hese chromosome-li$e vectors behave normally in mitosis and can carry more ()A than a !lasmid vector. Another advantage of eu$aryotic hosts is that they are ca!able of !roviding the !osttranslational modifications that many !roteins require. o +uch modifications may include adding carbohydrates or li!ids.

Eukaryote genes can be expressed in bacterial host cells.

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=or some mammalian !roteins, the host must be an animal cell to !erform the necessary modifications.

2,!erimental techniques facilitate the entry of foreign ()A into eu$aryotic cells. o In electroporation) brief electrical !ulses create a tem!orary hole in the !lasma membrane through which ()A can enter. o Alternatively, scientists can in;ect ()A into individual cells using microsco!ically thin needles. .o get ()A into !lant cells, the soil bacterium Agrobacterium can be used. /nce inside the cell, the ()A is incor!orated into the cell5s ()A by natural genetic recombination. he polymerase chain reaction !"CR# ampli$ies DNA in vitro. ()A cloning in cells remains the best method for !re!aring large quantities of a !articular gene or other ()A sequence. Ahen the source of ()A is scanty or im!ure, however, the polymera e chain reaction &*C+' is quic$er and more selective. .his technique can quic$ly am!lify any !iece of ()A without using cells. B o o C>' can ma$e billions of co!ies of a targeted ()A segment in a few hours. .his !rocess is faster than cloning via recombinant bacteria. In fact, C>' is being used increasingly to ma$e enough of a s!ecific ()A fragment to insert it directly into a vector, s$i!!ing the ste!s of ma$ing and screening a library.

In C>', a three-ste! cycle*heating, cooling, and re!lication*brings about a chain reaction that !roduces an e,!onentially growing !o!ulation of identical ()A molecules. o .he reaction mi,ture is heated to denature 7se!arate8 the ()A strands. o .he mi,ture is cooled to allow annealing 7hydrogen bonding8 of short, single-stranded ()A !rimers com!lementary to sequences on o!!osite sides at each end of the target sequence. o A heat-stable ()A !olymerase e,tends the !rimers in the 5 & direction. If a standard ()A !olymerase were used, the !rotein would be denatured along with the ()A during the first heating ste!. .he $ey to easy C>' automation was the discovery of an unusual ()A !olymerase, isolated from a bacterium living in hot s!rings, that can withstand the heat needed to se!arate the ()A strands at the start of each cycle. Dust as im!ressive as the s!eed of C>' is its s!ecificity. .he $ey to this high s!ecificity is the !rimers, which hydrogen-bond only to sequences at o!!osite ends of the target segment. /nly minute amounts of ()A need be !resent in the starting material, as long as a few molecules contain the com!lete target sequence. o .he ()A can be in a !artially degraded state. "y the end of the third cycle, one-fourth of the molecules are identical to the target segment, with both strands the a!!ro!riate length. Aith each successive cycle, the number of target segment molecules of the correct length doubles, soon greatly outnumbering all other ()A molecules in the reaction.

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(es!ite its s!eed and s!ecificity, C>' am!lification cannot substitute for gene cloning in cells when large amounts of a gene are desired. o /ccasional errors during C>' re!lication im!ose limits on the number of good co!ies that can be made. o Ahen C>' is used to !rovide the s!ecific ()A fragment for cloning, the resulting clones are sequenced to select clones with error-free inserts. (evised in 19E5, C>' has had a ma;or im!act on biological research and technology. C>' has am!lified ()A from a variety of sources4 fragments of ancient ()A from a F , year-old fro%en woolly mammoth< ()A from foot!rints or tiny amounts of blood or semen found at the scenes of violent crimes< ()A from single embryonic cells for the ra!id !renatal diagnosis of genetic disorders< and ()A of viral genes from cells infected with -IG.

Concept 20.2 DNA technology allow ! to t!#y the e,!ence) expre ion) an# "!nction o" a gene.
/nce scientists have !re!ared homogeneous sam!les of ()A, each containing a large number of identical segments, they can as$ some interesting questions about s!ecific genes and their functions. o (oes the sequence of the hummingbird -globin gene code for a !rotein structure that can carry o,ygen more efficiently than its counter!art in less metabolically active s!eciesH o (oes a !articular human gene differ from !erson to !ersonH o Are certain alleles of that gene associated with a hereditary disorderH o Ahere in the body and when during develo!ment is a given gene e,!ressedH o Ahat role does a certain gene !lay in an organismH .o answer these questions, researchers need to $now the nucleotide sequence of the gene and its counter!arts in other individuals and s!ecies, as well as its e,!ression !attern. One method o$ rapidly analyzing and comparing genomes is gel electrophoresis. -el electrophore i se!arates macromolecules*nucleic acids or !roteins*on the basis of their rate of movement through a !olymer gel in an electrical field. o .he rate of movement of each molecule de!ends on its si%e, electrical charge, and other !hysical !ro!erties. In restriction fragment analysis, the ()A fragments !roduced by restriction en%yme digestion of a ()A molecule are sorted by gel electro!horesis. o Ahen the mi,ture of restriction fragments from a !articular ()A molecule undergoes electro!horesis, it yields a band !attern characteristic of the starting molecule and the restriction en%yme used. o .he relatively small ()A molecules of viruses and !lasmids can be identified sim!ly by their restriction fragment !atterns. .he se!arated fragments can be recovered undamaged from gels, !roviding !ure sam!les of individual fragments. +cientists can use restriction fragment analysis to com!are two different ()A molecules re!resenting, for e,am!le, different alleles of a gene. o "ecause the two alleles differ slightly in ()A sequence, they may differ in one or more restriction sites.

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If the alleles do differ in restriction sites, each !roduces different-si%ed fragments when digested by the same restriction en%yme. In gel electro!horesis, the restriction fragments from the two alleles !roduce different band !atterns, allowing researchers to distinguish the two alleles.

'estriction fragment analysis is sensitive enough to distinguish between two alleles of a gene that differ by only one base !air in a restriction site, such as the normal and sic$le-cell alleles of the -globin gene. A method called .o!thern blotting combines gel electro!horesis with nucleic acid hybridi%ation. o /ne of this method5s many a!!lications is to identify hetero%ygous carriers of mutant alleles associated with genetic disease, although more ra!id methods involving C>' am!lification are currently used for this. o "ecause gel electro!horesis yields too many bands to distinguish individually, scientists use nucleic acid hybridi%ation with a s!ecific !robe to label discrete bands that derive from the gene of interest. o .he !robe is a radioactive, single-stranded ()A molecule that is com!lementary to the gene of interest 7for e,am!le, the -globin gene8. he entire nucleotide se%uence o$ a gene can be determined. 3ene sequencing has been automated based on a technique called the dideo yribonucleotide 7or dideo y, for short8 chain termination method. Inowing the entire nucleotide sequence of a gene allows researchers to com!are it to genes in other s!ecies, whose function may be $nown. If two genes from different s!ecies are similar in sequence, their gene !roducts li$ely !erform similar functions. Other experimental approaches analyze &hen and &here a gene is expressed. Jabeled nucleic acid !robes that hybridi%e with m')As can !rovide information about the time or !lace in the organism at which a gene is transcribed. =or e,am!le, there are two ways to find out how the e,!ression of the -globin gene changes during the embryonic develo!ment of the hummingbird. In the first method, called Northern blotting, scientists carry out gel electro!horesis on sam!les of m')A from hummingbird embryos at different stages of develo!ment, transfer the sam!les to a nitrocellulose membrane, and then allow the m')As on the membrane to hybridi%e with a labeled !robe that recogni%es -globin m')A. o If the m')A band is seen at a !articular stage, scientists can hy!othesi%e that the !rotein functions during events ta$ing !lace at that stage. A new and more sensitive method is called the rever e tran cripta e/polymera e chain reaction, or +T/*C+. o Analysis of hummingbird -globin gene e,!ression with '.-C>' begins similarly to )orthern blotting, with the isolation of m')As from different develo!mental stages of hummingbird embryos. o )e,t, reverse transcri!tase is added to ma$e c()A, which then serves as a tem!late for C>' am!lification using !rimers from the -globin gene. o Ahen the !roducts are run on a gel, co!ies of the am!lified region are observed as bands only in sam!les that originally contained the -globin m')A.

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In the case of hummingbird -globin, scientists might e,!ect to see a band a!!ear at the stage when red blood cells begin forming, with all subsequent stages showing the same band. '.-C>' can also be carried out with m')As collected from different tissues at one time to discover which tissue is !roducing a s!ecific m')A. o +!ecific m')As can be identified by using labeled !robes in !lace, or in situ, in the intact organism. o .his technique, called in situ hybri#i$ation, is most often carried out with !robes labeled by the attachment of fluorescent dyes. Ahen the entire genomes of a number of organisms have been sequenced, it is !ossible to study the e,!ression of large grou!s of genes, to study how genes act together to !roduce and maintain a functioning organism. 'esearchers use genome sequences as !robes to investigate which genes are transcribed in different situations, such as in different tissues or at different stages of develo!ment. 'esearchers also loo$ for grou!s of genes that are e,!ressed in a coordinated manner, with the aim of identifying networ$s of gene e,!ression across an entire genome. .he basic strategy in such global 7genome-wide8 e,!ression studies is to isolate the m')As made in !articular cells, use these molecules as tem!lates for ma$ing the corres!onding c()As by reverse transcri!tion, and then em!loy nucleic acid hybridi%ation to com!are this set of c()As with a collection of ()A fragments re!resenting all or !art of the genome. .he results identify the subset of genes in the genome that are being e,!ressed at a given time or under certain conditions. 3enome-wide e,!ression studies are made !ossible by the use of DNA microarray a ay . o A ()A microarray consists of tiny amounts of a large number of single-stranded ()A fragments re!resenting different genes fi,ed to a glass slide in a tightly s!aced array, or grid, also called a DNA chip. o Ideally, these fragments re!resent all the genes of an organism. o .he ()A fragments on a microarray are tested for hybridi%ation with c()A molecules that have been !re!ared from the m')As in !articular cells of interest and labeled with fluorescent dyes. In one e,am!le of a global e,!ression study, researchers !erformed ()A microarray assays on more than 9 K of the genes of the nematode !aenorhabditis elegans during every stage of its life cycle. .he e,!ression of nearly L K of the genes changed dramatically during develo!ment. 6any genes were e,!ressed in a se,-s!ecific !attern. In addition to uncovering gene interactions and !roviding clues to gene function, ()A microarray assays may contribute to a better understanding of certain diseases and suggest new diagnostic techniques or thera!ies. o >om!aring !atterns of gene e,!ression in breast cancer tumors and noncancerous breast tissue, for e,am!le, has already resulted in more informed and effective treatment !rotocols. 'cientists disable genes and obser(e the conse%uences to determine the $unction o$ the genes.

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In an a!!lication called in (itro m!tagene i , s!ecific mutations are introduced into the sequence of a cloned gene, and then the mutated gene is returned to a cell in such a way that it disables 70$noc$s out18 the normal cellular co!ies of the same gene. If the introduced mutations alter or destroy the function of the gene !roduct, the !henoty!e of the mutant cell may hel! reveal the function of the missing normal !rotein. 'esearchers can even !ut such a mutated gene into cells from the early embryo of a multicellular organism 7such as a mouse8 to study the role of the gene in the develo!ment and functioning of the whole organism. In 2 L, government agencies in the Mnited +tates, >anada, >hina, and the 2uro!ean Mnion announced several large-scale !ro;ects whose aim is to disable every one of the 25, genes in the mouse genome. A newer method for silencing the e,!ression of selected genes e,!loits the !henomenon of +NA inter"erence &+NAi'. .his a!!roach uses synthetic, double-stranded ')A molecules that match the sequence of a !articular gene to trigger the brea$down of the gene5s messenger ')A or to bloc$ its translation. o .o date, the ')Ai technique has been used successfully to reduce 70$noc$ down18 the e,!ression of s!ecific genes in mammalian cells, including human cells in culture, and !lans are under way to try using ')Ai for the treatment of human disorders such as macular degeneration of the eye. o In one study, ')Ai was used to !revent the e,!ression of ELK of the genes in early nematode embryos, one gene at a time. o Analysis of the !henoty!es of the worms that develo!ed from these embryos enabled the researchers to classify most of the genes into a small number of grou!s by function.

Concept 20.0 Cloning organi m may lea# to the pro#!ction o" tem cell "or re earch an# other application .
+cientists can now clone multicellular organisms from single cells. o .his is called organismal cloning to distinguish it from gene cloning and cell cloning, the division of an ase,ually re!roducing cell into a collection of genetically identical cells. /rganismal cloning has the !otential to generate stem cells, which can develo! into many different tissues. .he cloning of !lants and animals was first attem!ted more than 5 years ago in e,!eriments designed to determine whether all the cells of an organism have the same genes 7a conce!t called genomic e"uivalence8 or whether cells lose genes during the !rocess of differentiation. )hole plants ha(e been cloned $rom single di$$erentiated cells since the *+,-s. +uccessful attem!ts to clone whole !lants from single differentiated cells were made during the 195 s by =. >. +teward of >ornell Mniversity. +teward and his students found that cultured, differentiated root cells could grow into normal adult !lants, each genetically identical to the !arent !lant, thus demonstrating that differentiation does not necessarily involve irreversible changes in the ()A. In !lants, mature cells can dedifferentiate and then give rise to all the s!eciali%ed cell ty!es of the organism. Any cell with this !otential is said to be totipotent.

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Clant cloning is now used e,tensively in agriculture to re!roduce !lants that have valuable characteristics, such as the ability to resist a !lant !athogen. Nuclear transplantation &as used in animal experiments. (ifferentiated cells from animals do not readily divide in culture. 2arly researchers investigating whether differentiated animal cells can be toti!otent removed the nucleus of an unfertili%ed frog5s egg or %ygote and re!laced it with the nucleus of a differentiated cell from a tad!ole, a !rocedure called nuclear transplantation. If the nucleus from the differentiated donor cell retains its full genetic ca!ability, then it should be able to direct develo!ment of the reci!ient egg into all the tissues and organs of an organism. .he trans!lanted nucleus was often able to su!!ort normal develo!ment of the egg into a tad!ole. -owever, the !otential of a trans!lanted nucleus to direct normal develo!ment was inversely related to the age of the donor4 .he older the donor nucleus, the lower the !ercentage of normally develo!ing tad!oles. >learly, something in the nucleus changed as the tad!ole5s cells differentiated. o In frogs and most other animals, nuclear !otential tends to be increasingly restricted as embryonic develo!ment and cell differentiation !rogress. In 199#, +cottish researchers ca!tured news!a!er headlines when they announced the birth of (olly, a lamb cloned from an adult shee! by nuclear trans!lantation from a differentiated cell. .hese researchers cultured donor mammary cells in a nutrient-!oor medium and then fused these cells with enucleated shee! eggs. .he resulting di!loid cells divided to form early embryos, which were im!lanted into surrogate mothers. o /ut of several hundred im!lanted embryos, one successfully com!leted normal develo!ment, and (olly was born. Jater analyses showed that (olly5s chromosomal ()A was identical to that of the nucleus donor. (olly5s mitochondrial ()A came from the egg donor. In 2 &, at age L, (olly suffered com!lications from a lung disease usually seen in only much older shee!, and she was euthani%ed. (olly5s !remature death led to s!eculation that her cells were not quite as 0healthy1 as those of a normal shee!, !ossibly reflecting incom!lete re!rogramming of the original trans!lanted nucleus. +ince 199#, reproductive cloning*the !roduction of new individuals*has been demonstrated in many other mammals, including mice, cats, cows, horses, mules, !igs, and dogs. >loned animals of the same s!ecies do not always loo$ or behave identically. o In a herd of cows cloned from the same line of cultured cells, certain cows are dominant and others are more submissive. o .he first cloned cat, named >> for >arbon >o!y has a calico coat, li$e her single female !arent, but the color and !attern are different because of random : chromosome inactivation, which is a normal occurrence during embryonic develo!ment.
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Lecture Outline for Campbell/Reece Biology, 8th Edition, Pearson Education, Inc.

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Identical human twins, naturally occurring 0clones,1 always differ somewhat. 2nvironmental influences and random !henomena can !lay a significant role during develo!ment.

.he successful cloning of so many mammals has led to s!eculation about the cloning of humans. )uclei from differentiated human cells have been trans!lanted into unfertili%ed enucleated eggs, and the eggs stimulated to divide. In 2 1, a research grou! in 6assachusetts observed a few early cell divisions in such an e,!eriment. here are problems associated &ith animal cloning. In most nuclear trans!lantation studies, very few cloned embryos develo! normally to birth and many of those e,hibit defects. o 2ven cloned animals that a!!ear normal li$ely have subtle defects. In the nuclei of fully differentiated cells, the e,!ression of many genes is re!ressed as a result of e!igenetic changes in chromatin, such as acetylation of histones or methylation of ()A. (uring the nuclear transfer !rocedure, many of these changes must be reversed in the laterstage nucleus from a donor animal in order for genes to be e,!ressed or re!ressed a!!ro!riately in early stages of develo!ment. ()A in embryonic cells from cloned embryos, li$e ()A of differentiated cells, tends to have more methyl grou!s than ()A in equivalent cells from uncloned embryos of the same s!ecies. .his suggests that the re!rogramming of donor nuclei requires chromatin restructuring, which does not occur fully during cloning !rocedures. "ecause ()A methylation hel!s regulate gene e,!ression, mis!laced methyl grou!s in the ()A of donor nuclei may interfere with the !attern of gene e,!ression necessary for normal embryonic develo!ment. 'tem cells can be used to treat human diseases. +cientists5 ma;or aim in cloning human embryos is the !roduction of stem cells for treating human diseases. A tem cell is a relatively uns!eciali%ed cell that can both re!roduce itself indefinitely and, under a!!ro!riate conditions, differentiate into s!eciali%ed cells of one or more ty!es. .hus, stem cells are able both to re!lenish their own !o!ulation and to generate cells that travel down s!ecific differentiation !athways. 6any early animal embryos contain stem cells ca!able of giving rise to any ty!e of differentiated embryonic cells. o Embryonic stem cells can be isolated from early embryos at the blastula or blastocyst stage. .he adult body also has adult stem cells, which are able to give rise to many, but not all, cell ty!es. o =or e,am!le, bone marrow contains several ty!es of stem cells, including one that can generate all the different $inds of blood cells and another that can differentiate into bone, cartilage, fat, muscle, and the linings of blood vessels. o 2ven the adult brain contains stem cells that !roduce certain $inds of nerve cells.

Lecture Outline for Campbell/Reece Biology, 8th Edition, Pearson Education, Inc.

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+cientists are learning to identify and isolate stem cells from various tissues and, in some cases, to grow them in culture. Aith the addition of s!ecific growth factors, cultured stem cells from adult animals have been made to differentiate into multi!le ty!es of s!eciali%ed cells. .his technology may !rovide cells for the re!air of damaged or diseased organs, such as insulin-!roducing !ancreatic cells for !eo!le with diabetes or certain $inds of brain cells for !eo!le with Car$inson5s disease or -untington5s disease. 2mbryonic stem cells offer more !otential than adult stem cells for medical a!!lications, at least for now. -owever, deriving embryonic stem cells from human embryos raises ethical and !olitical issues. B 2mbryonic stem cells are currently obtained from embryos donated by !atients undergoing infertility treatment or from long-term cell cultures originally established from donated embryos. B If scientists can clone human embryos to the blastocyst stage, they might be able to use such clones as the source of embryonic stem cells. o A donor nucleus from a !atient with a !articular disease could allow the !roduction of embryonic stem cells for treatment that match the !atient and are not re;ected by his or her immune system. Although most !eo!le believe that the re!roductive cloning of humans is unethical, o!inions vary about the morality of therapeutic cloning, whose ma;or aim is to !roduce embryonic stem cells to treat disease. +ome believe it is wrong to create embryos that will be destroyed, whereas others, in the words of the researcher who created (olly, believe that 0cloning !romises such great benefits that it would be immoral not to do it.1

Concept 20.1 The practical application o" DNA technology a""ect o!r live in many way .
DNA technology is reshaping medicine and the pharmaceutical industry. 6odern biotechnology is ma$ing enormous contributions both to the diagnosis of diseases and in the develo!ment of !harmaceutical !roducts. .he identification of genes whose mutations are res!onsible for genetic diseases may lead to ways to diagnose, treat, or even !revent these conditions. +usce!tibility to many 0nongenetic1 diseases, from arthritis to AI(+, is influenced by a !erson5s genes. o (iseases of all sorts involve changes in gene e,!ression within the affected genes and within the !atient5s immune system. o Msing ()A microarray assays and other techniques to com!are gene e,!ressions in healthy and diseased tissue can identify these changes and lead to the develo!ment of targets for !revention or thera!y. C>' and labeled nucleic acid !robes can trac$ down the !athogens res!onsible for infectious diseases. o =or e,am!le, '.-C>' can am!lify and thus detect -IG ()A in blood and tissue sam!les, detecting an otherwise elusive infection.
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Lecture Outline for Campbell/Reece Biology, 8th Edition, Pearson Education, Inc.

6edical scientists can use ()A technology to identify individuals who have genetic diseases before the onset of sym!toms, even before birth. 3enetic disorders are diagnosed by using C>' and !rimers corres!onding to cloned disease genes, and then sequencing the am!lified !roduct to loo$ for the disease-causing mutation. o >loned disease genes include those for sic$le-cell disease, hemo!hilia, cystic fibrosis, -untington5s disease, and (uchenne muscular dystro!hy. It is !ossible to detect abnormal allelic forms of genes even in cases in which the gene has not yet been cloned. .he !resence of an abnormal allele can be diagnosed with reasonable accuracy if a closely lin$ed genetic mar#er has been found. o A genetic mar$er is a ()A sequence that varies in a !o!ulation< in a gene, such sequence variation is the basis of different alleles. Dust li$e coding sequences, noncoding ()A at a s!ecific !lace or locus on a chromosome may e,hibit small nucleotide differences among individuals, or polymorphisms. +ingle base-!air variations in the genomes of the human !o!ulation serve as useful genetic mar$ers. A single base-!air site where variation is found in at least 1K of the !o!ulation is called a ingle n!cleoti#e polymorphi m 7.N*8. o +)Cs occur on average about once in 1 to & base !airs in the human genome, in both coding and noncoding sequences. +ome +)Cs alter the sequence recogni%ed by a restriction en%yme, changing the lengths of the restriction fragments formed by digestion with that en%yme. o .his ty!e of sequence change is called a re triction "ragment length polymorphi m 7+23*, !ronounced 0'if-li!18. .oday, +)Cs can be detected by very sensitive microarray analysis or by C>'. .he !resence of an abnormal allele that causes a genetic disorder can be diagnosed with reasonable accuracy if a closely lin$ed +)C mar$er has been found. o Alleles for -untington5s disease and a number of other genetic diseases were first detected by means of '=JCs in this indirect way. o If the mar$er and the gene itself are close enough, crossing over between the mar$er and the gene is very unli$ely to occur during gamete formation, and the two regions are almost always inherited together. .uman gene therapy holds great potential. -ene therapy*introducing genes into an afflicted individual for thera!eutic !ur!oses* holds great !otential for treating genetic disorders caused by a single gene. In theory, a normal allele of the defective gene could be inserted into dividing somatic cells of the tissue affected by the disorder, such as bone marrow cells. o "one marrow cells, which include the stem cells that give rise to all the cells of the blood and immune system, are !rime candidates for gene thera!y. +evere combined immunodeficiency 7+>I(8, caused by a single defective gene, has been treated in gene thera!y trials. o If successful, gene thera!y cures the !atient, whose bone marrow cells then !roduce the missing !rotein. o In 2 , ten young children with +>I( were treated by gene thera!y.
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After two years, nine of these !atients showed significant im!rovement. -owever, three of the !atients subsequently develo!ed leu$emia and one died. .he retroviral vector used to carry the normal allele into bone marrow cells had inserted near a gene involved in the !roliferation and develo!ment of blood cells, which may have caused leu$emia. In a recent e,!eriment on mice, a re!lacement of the mouse version of the same gene led to a high incidence of lym!homa, a blood cancer. Cerha!s an un$nown function of the gene itself may be res!onsible.

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3ene thera!y also raises technical questions. o -ow can the activity of the transferred gene be controlled so that cells ma$e a!!ro!riate amounts of the gene !roduct at the right time and in the right !laceH o -ow can scientists be sure that the insertion of the thera!eutic gene does not harm some other necessary cell functionH In addition to these technical challenges, gene thera!y raises ethical questions. +ome critics believe that tam!ering with human genes in any way is immoral. /thers see no difference between the trans!lantation of genes into somatic cells and the trans!lantation of organs. .he treatment of human germ-line cells in the ho!e of correcting a defect in future generations raises ethical questions. o Mnder what circumstances, if any, should we alter the genomes of human germ linesH o Aould this inevitably lead to the !ractice of eugenics, a deliberate effort to control the genetic ma$eu! of human !o!ulationsH =rom a biological !ers!ective, the elimination of unwanted alleles from the gene !ool could be !roblematic because genetic variation is a necessary ingredient for the survival of a s!ecies as environmental conditions change with time. o 3enes that are damaging under some conditions may be advantageous under other conditions. o (o we have the right to ma$e genetic changes that could be detrimental to the survival of our s!ecies in the futureH he pharmaceutical industry uses biotechnology to de(elop ne& drugs. +mall molecules can be tailored to combat certain cancers by bloc$ing the function of a !rotein crucial for the tumor cells5 survival. /ne drug, called 3leevec 7or imatinib8, is a small molecule that inhibits a s!ecific rece!tor tyrosine $inase. .he overe,!ression of this rece!tor, resulting from a chromosomal translocation, is instrumental in causing chronic myelogenous leu$emia 7>6J8. Catients in the early stages of >6J who have been treated with 3leevec have e,hibited nearly com!lete, sustained remission from the cancer. +imilar drugs have also been used to treat a few ty!es of lung and breast cancers, whose molecular basis is fairly well understood. "roteins are produced in cell cultures. ()A cloning and gene e,!ression systems can !roduce large quantities of !roteins that are !resent naturally in only minute amounts.
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Lecture Outline for Campbell/Reece Biology, 8th Edition, Pearson Education, Inc.

.he host cells used in these e,!ression systems can be engineered to secrete a !rotein as it is made, thus removing the need for its !urification. Among the first !harmaceutical !roducts 0manufactured1 in this way were human insulin and human growth hormone 7-3-8. o +ome 2 million !eo!le with diabetes in the Mnited +tates require daily insulin in;ections. o -uman growth hormone has been used to treat children born with a form of dwarfism caused by inadequate amounts of -3-. Another im!ortant !harmaceutical !roduct created by genetic engineering is tissue !lasminogen activator 7.CA8. o If administered shortly after a heart attac$, .CA hel!s dissolve blood clots and reduces the ris$ of subsequent heart attac$s. >ells in culture can also be used to !roduce vaccines, which stimulate the immune system to defend against s!ecific !athogens. Alternatively, genetic engineering can be used to modify a !athogen5s genome, resulting in a wea$ened !athogen that can then serve as a vaccine. "roteins are produced by /pharm0 animals and plants. In some cases, whole animals are used to !roduce !roteins, instead of cell systems. Msing e,!erimental methods, scientists introduce a gene from one animal into the genome of another animal, which is then called a tran genic animal. .o do this, scientists first remove eggs from a female and fertili%e them in vitro. .hey then in;ect the cloned ()A from another organism directly into the nuclei of the fertili%ed eggs. +ome of the cells integrate the foreign ()A, the transgene, into their genomes and are able to e,!ress the foreign gene. .he engineered embryos are then surgically im!lanted into a surrogate mother. If an embryo develo!s successfully, the result is a transgenic animal, containing a gene from a third 0!arent.1 Assuming the introduced gene encodes a !rotein desired in large quantities, these transgenic animals can act as !harmaceutical 0factories.1 o =or e,am!le, a transgene for a human blood !rotein such as antithrombin has been inserted into the genome of a goat so that antithrombin is secreted in the animal5s mil$. o .he !rotein can then be !urified from the mil$. o 'esearchers have also engineered transgenic chic$ens that e,!ress large amounts of the transgene5s !roduct in eggs.

.he human !roteins !roduced by farm animals may differ in some ways from the corres!onding natural human !roteins, so they must be tested carefully to ensure that they will not cause allergic reactions or other adverse effects in !atients who receive them. In a novel twist, the !harmaceutical industry is beginning to develo! 0!harm1 !lants, analogous to 0!harm1 animals. 0Charm1 !lants ma$e human !roteins for medical use and viral !roteins for use as vaccines. o In 2 #, the M.+. =ood and (rug Administration a!!roved a !lan to !lant more than &, acres in Iansas with rice containing genes for mil$ !roteins, used to treat infant diarrhea.

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1enetic pro$iles may be important $orensic e(idence. "ody fluids or small !ieces of tissue may be left at the scene of a violent crime or on the clothes of the victim or assailant. If enough blood, semen, or tissue is available, forensic laboratories can determine the blood ty!e or tissue ty!e by using antibodies to detect s!ecific cell-surface !roteins. +uch tests require fairly fresh sam!les in relatively large amounts, however, and can serve only to e,clude a !articular sus!ect. ()A testing, in contrast, can identify the guilty individual with a high degree of certainty because the ()A sequence of every !erson is unique 7e,ce!t for identical twins8. 3enetic mar$ers that vary in the !o!ulation can be analy%ed for a given !erson to determine that individual5s unique set of genetic mar$ers, or genetic pro"ile. .he ="I started a!!lying ()A technology in forensics in 19EE, using '=JC analysis by +outhern blotting to detect similarities and differences in ()A sam!les. o .his method required much smaller sam!les of blood or tissue than earlier methods* only about 1, cells. .oday, in !lace of '=JCs, forensic scientists usually use an even more sensitive method, which ta$es advantage of genetic mar$ers called hort tan#em repeat &.T+ '. +.'s are variations in the lengths of certain re!eated base sequences in s!ecific regions of the genome. .he number of re!eats !resent in these regions is highly !olymor!hic, varying from !erson to !erson. o =or e,am!le, one individual may have the sequence A>A. re!eated & times at one genome locus, whereas another individual may have 1E re!eats at this locus. C>' is used to am!lify !articular +.'s using sets of !rimers that are labeled with different colored fluorescent tags< the length of the region, and thus the number of re!eats, can then be determined by electro!horesis. o +ince +outhern blotting is not required, this method is quic$er than '=JC analysis. .he C>' ste! allows the method to be used even when the ()A is in !oor condition or available in only minute quantities. o A tissue sam!le containing as few as 2 cells can be sufficient for C>' am!lification. o In a murder case, for e,am!le, this method can be used to com!are ()A sam!les from the sus!ect, the victim, and a small amount of blood found at the crime scene. =orensic scientists test only a few selected !ortions of the ()A*about 1& +.' mar$ers. .he !robability that two !eo!le 7who are not identical twins8 would have e,actly the same set of +.' mar$ers is vanishingly small. .he Innocence Cro;ect, a non!rofit organi%ation dedicated to overturning wrongful convictions, uses +.' analysis of archived sam!les from crime scenes to revisit old cases. o As of 2 L, 1E innocent !eo!le had been released from !rison as a result of forensic and legal wor$ by this grou!. In another e,am!le of genetic !rofiling, a com!arison of the ()A of a mother, her child, and the !ur!orted father can conclusively settle a question of !aternity. B +ometimes !aternity is of historical interest4 3enetic !rofiles !rovided strong evidence that .homas Defferson or one of his close male relatives fathered at least one of the children of his slave, +ally -emings.
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Lecture Outline for Campbell/Reece Biology, 8th Edition, Pearson Education, Inc.

Analy%ing genetic !rofiles can also identify victims of mass casualties. o After the Aorld .rade >enter attac$ in 2 1, more than 1 , sam!les of victims5 remains were com!ared with ()A traces on !ersonal items !rovided by families. o .hese com!arisons led to the identification of nearly &, victims. Dust how reliable is a genetic !rofileH .he greater the number of mar$ers e,amined in a ()A sam!le, the more li$ely it is that the !rofile is unique to one individual. In forensic cases using +.' analysis with 1& mar$ers, the !robability of two !eo!le having identical ()A !rofiles is somewhere between one chance in 1 billion and one in several trillion. .he e,act !robability de!ends on the frequency of those mar$ers in the general !o!ulation. o Information on how common various mar$ers are in different ethnic grou!s is critical because these mar$er frequencies may vary considerably among ethnic grou!s and between a !articular ethnic grou! and the !o!ulation as a whole. Aith the increasing availability of frequency data, forensic scientists can ma$e e,tremely accurate statistical calculations. 3enetic !rofiles are now acce!ted as com!elling evidence by legal e,!erts and scientists ali$e. 2icroorganisms can be used $or en(ironmental cleanup. As a grou!, microorganisms have a remar$able ability to transform chemicals. 6any bacteria can e,tract heavy metals, such as co!!er, lead, and nic$el, from their environments and incor!orate the metals into com!ounds such as co!!er sulfate or lead sulfate, which are readily recoverable. 3enetically engineered microbes may become im!ortant in both mining minerals 7es!ecially as ore reserves are de!leted8 and cleaning u! highly to,ic mining wastes. "iotechnologists are also trying to engineer microbes that can be used in wastewater treatment !lants to degrade chlorinated hydrocarbon. 3io$uels can supplement or replace $ossil $uels. "iofuels from cro!s such as corn, soybeans, and cassava have been !ro!osed as a su!!lement or even re!lacement for fossil fuels. o .o !roduce 0bioethanol,1 starch is converted to sugar and then fermented by microorganisms< a similar !rocess can !roduce 0biodiesel1 from !lant oils. 2ither !roduct can be mi,ed with gasoline or used alone to !ower vehicles. >ritics charge that the environmental effects of growing these cro!s ma$es their total im!act greater than that of fossil fuels. DNA technology has agricultural applications. .he selective breeding of livestoc$, or animal husbandry, has e,!loited naturally occurring mutations and genetic recombination for centuries. ()A technology !ermits the genetic engineering of transgenic animals, which s!eeds u! the selective breeding !rocess. o .he goals of creating a transgenic animal are often the same as the goals of traditional breeding*for instance, to ma$e a shee! with better quality wool, a !ig with leaner meat, or a cow that will mature in a shorter time.
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+cientists might, for e,am!le, identify and clone a gene that leads to the develo!ment of larger muscles 7muscles ma$e u! most of the meat we eat8 in one variety of cattle and transfer it to other cattle or even to shee!. .his mani!ulation may cause low fertility or increased susce!tibility to disease, however.

Clants are easier to genetically engineer than most animals because a single tissue cell grown in culture and genetically mani!ulated can give rise to an adult !lant with new traits. .he most commonly used vector for introducing new genes into !lant cells is a !lasmid, called the Ti pla mi#, from the soil bacterium Agrobacterium tumefaciens. o .his !lasmid integrates a segment of its ()A, $nown as . ()A, into the chromosomal ()A of its host !lant cells. o =or vector !ur!oses, researchers wor$ with versions of the !lasmid that do not cause disease. +cientists can introduce recombinant .i !lasmids into !lant cells by electro!oration or by infecting !lant cells in culture with bacteria containing the recombinant !lasmid. Alternatively, the recombinant !lasmid can be !ut bac$ into Agrobacterium< susce!tible !lants or !lant cells growing in culture are then infected with bacteria that contain the recombinant !lasmid. 3enetic engineering is ra!idly re!lacing traditional !lant-breeding !rograms for sim!le genetic traits such as herbicide or !est resistance. B 3enetically engineered cro!s that can resist destructive insects have reduced the need for chemical insecticides.

In India, the insertion of a salinity-resistance gene from a coastal mangrove !lant into the genomes of several rice varieties has resulted in rice !lants that can grow in water three times as salty as seawater. o .he research foundation that carried out this genetic engineering estimates that one-third of all irrigated land has high salinity owing to over-irrigation and intensive use of chemical fertili%ers, which re!resents a serious threat to the food su!!ly. +alinityresistant cro! !lants would be enormously valuable worldwide. 3enetic engineering also has great !otential for im!roving the nutritional value of cro! !lants. o =or instance, scientists have develo!ed transgenic rice !lants that !roduce yellow rice grains containing beta-carotene, which our body uses to ma$e vitamin A. o .his 0golden1 rice could hel! !revent vitamin A deficiency in the half of the world5s !o!ulation that de!ends on rice as a sta!le food. DNA technology raises some sa$ety and ethical concerns. 2arly concerns about !otential dangers associated with recombinant ()A technology focused on the !ossibility that ha%ardous new !athogens might be created. o Ahat might ha!!en, for instance, if cancer cell genes were transferred into bacteria or virusesH .o guard against such rogue microbes, scientists develo!ed strict laboratory !rocedures designed to !rotect researchers from infection by engineered microbes and to !revent the microbes from accidentally leaving the laboratory. +trains of microorganisms to be used in recombinant ()A e,!eriments are genetically cri!!led to ensure that they cannot survive outside the laboratory, and certain obviously dangerous e,!eriments have been banned.
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Lecture Outline for Campbell/Reece Biology, 8th Edition, Pearson Education, Inc.

.oday, most !ublic concern about !ossible ha%ards centers not on recombinant microbes but on genetically mo#i"ie# &-4' organisms used as food. A 36 organism is one that has acquired by artificial means one or more genes from another s!ecies or even from another variety of the same s!ecies. o +almon, for e,am!le, have been genetically modified by the addition of a more active salmon growth hormone gene. 36 cro!s are wides!read in the Mnited +tates, Argentina, and "ra%il< together these countries account for more than E K of the world5s acreage devoted to 36 cro!s. o In the Mnited +tates, the ma;ority of corn, soybeans, and canola are 36 cro!s, and 36 !roducts are not required to be labeled. 36 cro!s are a sub;ect of ongoing controversy in 2uro!e, where the 36 revolution has met with strong o!!osition from countries concerned about the safety of 36 foods and !ossible environmental consequences of growing 36 !lants. B Although a small number of 36 cro!s have been grown on 2uro!ean soil, their !roducts have generally failed in local mar$ets, and the future of 36 cro!s in 2uro!e is uncertain. 2arly in 2 , negotiators from 1& countries 7including the Mnited +tates8 agreed on a "iosafety Crotocol that requires e,!orters to identify 36 organisms !resent in bul$ food shi!ments and allows im!orting countries to decide whether the !roducts !ose environmental or health ris$s. Advocates of a cautious a!!roach toward 36 cro!s fear that transgenic !lants might !ass their new genes on to close relatives in nearby wild areas. o If cro! !lants carrying genes for resistance to herbicides, diseases, or insect !ests !ollinated wild !lants, the offs!ring might become 0su!er weeds1 that are very difficult to control. Another !ossible ha%ard, suggested by one laboratory-based study, is that a transgene encoding a !esticide-ty!e !rotein might cause !lants to !roduce !ollen to,ic to butterflies. -owever, scientists with the Agricultural 'esearch +ervice concluded from a two-year study that butterflies are unli$ely to be e,!osed to to,ic levels of !ollen. +ome !eo!le fear that the !rotein !roducts of transgenes might lead to allergic reactions, although advocates suggest that advance tests would !revent the !roduction of such !roteins. In the Mnited +tates, new a!!lications of biotechnology are evaluated for !otential ris$s by various regulatory agencies. Advances in biotechnology have enabled scientists to obtain com!lete genome sequences for humans and many other s!ecies, !roviding a vast treasure trove of information about genes. .he increasing s!eed and decreasing cost with which genome sequences of individual !eo!le can be determined are raising significant ethical questions. o Aho should have the right to e,amine someone else5s genetic informationH o -ow should a !erson5s genetic information be usedH o +hould a !erson5s genome be a factor in suitability for a ;ob or eligibility for insuranceH .he !ower of ()A technology and genetic engineering*our ability to !rofoundly and ra!idly alter s!ecies that have been evolving for millennia*demands that we !roceed with humility and caution.

Lecture Outline for Campbell/Reece Biology, 8th Edition, Pearson Education, Inc.

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