Tortora Chapter 6: Microbial Growth

The Requirements for Growth

Tow main categories:
>physical
- temp., pH, osmotic pressure
>chemical
- sources of C, N, S, P, O, trace elements, and organic growth factors
Physical requirements
Temperature
>psychrophiles (cold-loving microbes)
>mesophiles (moderate-temperature-loving)
>thermophiles (heat-loving microbes)
-most bacteria grow only w/in limited range of temps
-their max & min growth temps are only about 30C apart
-they grow poorly at the high & low temp extremes w/in their range
>minimum growth temperature - lowest temp @ w/c the species will grow
>optimum growth temperature - temp @ w/c the species grows best
>maximum growth temperature - highest temp @ w/c growth is possible
-the reproductive rate drops off very quickly at temperatures only a little abov
e the optimum bc the high temp has inactivated necessary enzymatic systems of th
e cell
>psychrophiles vs psychrotrophs
psychrophiles optimum temp = 15C; does not grow at 25C
psychotrophs optimum temp = 20-30C; does not grow at 40C; grow fairly
well at refrigerator temps; able to slowly degrade food
>mesophiles
optimum temp = 25-40C
optimum temp for many pathogenic bacteria is 37C
include most of the common spoilage and disease organisms
>thermophiles
optimum temp = 50-60C
sunlit soil, thermal waters like hot springs
endospores formed by thermophilic bacteria are unusally heat resistant
important in organic compost piles
>hyperthermophiles or extreme thermophiles
members of the Archaea
optimum temp = 80C or higher
hot springs in volcanic activity
sulfur is usually important in their metabolic activity
pH
-most bacteria grow best b/n pH 6.5-7.5
-a few grow at an acidic pH below pH 4 (acidophiles)
sauerkraut, pickles, many cheeses are preserved from spoilage by acids
produced by bacterial fermentation
molds & yeasts grow over a greater pH range than bacteria will, but the
optimum pH is generally below that of bacteria (pH 5-6)
-alkalinity also inhibits microbial growth but is rarely used to preserve foods
-when bacteria are cultured in vitro, their own acids might interfere with their
own growths -> needs buffer with peptones, phosphate salts, etc
Osmotic Pressure
-microbes require water for growth, their comp. is about 80-90% water
-in a hypertonic solution -> plasmolysis (shrinkage of cell cytoplasm) -> cell g
rowth is inhibited as the PM pulls away from the cell wall
-addition of salts for food preservation
-extreme halophiles: may be termed obligate halophiles bc they have adapted well
to high salt conc. that they actually require it for growth; those from the Dea

d Sea
-facultative halophiles: do not require high salt conc. but able to grow at salt
conc. up to 2% (conc. inhibits growth of most org.)
-most microbes must be grown in a medium that is nearly all water; agar (complex
polysaccharide isolated from marine algae) = 1.5%
Chemical Requirements
Water
Carbon
N, S, P
>nitrogen fixation: some bacteria, including photosynthesizing cyanobacteria, us
e gaseous nitrogen (N2) directly from the atmosphere
Trace elements - iron, copper, molybdenum, zinc (tap water)
Oxygen
>obligate aerobes: require oxygen to live
>facultative anaerobes: can use oxygen when it is present but are able to contin
ue growth by using fermentation of anaerobic respiration when O2 is not availabl
e; efficiency decreases in the absence of oxygen; Escherichia coli in GIT, many
yeasts
>obligate anaerobes: unable to use O2 for energy-yielding reactions; most are ha
rmed by it; genus Clostridium (cause tetanus & botulism)
>aerotolerant anaerobes: cannot use O2 for growth but can tolerate it well; many
of the aerotolerant bacteria characteristically ferment carbohydrates to lactic
acid, example of lactic acid-producing aerotolerant anaerobes is lactobacilli u
sed in production of many acidic fermented foods like pickles and cheese
>microaerophiles: require O2 conc. lower than those in air
Organic Growth Factors - essential organic compounds an organism is unable to sy
nthesize; must be directly obtained from the environment
Culture Media
>nutrient material prepared for the growth of microorganisms in a laboratory
>inoculum: microbes introduced into a culture medium to initiate growth
>culture: microbes that grow & multiply in/on a culture medium
>criteria for culture medium:
1. it must contain the right nutrients, sufficient moisture, properly
adjusted pH, suitable level of O2
2. it must initially be sterile (contains no living microorganisms)
3. should be incubated at the proper temp.
>agar
-complex polysaccharide derived from a marine alga
-solidifying agent for solid medium
-few microbes can degrade agar, so it remains solid
-liquefies at around 100C; in lab, it is held in water baths @ 50C
-slants: at an angle in a test tube
-deep: solidified in a vertical tube
-Petri plates
Chemically Defined Media
>one whose exact chem comp. is known
>E. coli, a chemoheterotroph, needs glucose in the medium
>fastidious: organisms that require many growth factors; example is Lactobacillu
s; used in tests that determine the conc. of a particular vitamin in a substance
(amt of lactic acid produced proportional to the amt of vitamin in test substan
ce)
Complex Media
>most heterotrophic bacteria and fungi are routinely grown on complex media
>made up of nutrients including extracts from yeasts, meat, plants, digests of p
roteins, etc
>exact chem. comp. varies slightly from batch to batch

>the energy, C, N, S requirements of the microbe are provided primarily by prote

in -> reduced to shorter chains of AA called peptones, can be digested by bacter
ia
>vitamins & other OGFs are provided by meat extracts or yeast extracts; yeast ex
tracts particularly rich in B vitamins
>nutrient broth: complex medium in liquid form
>nutrient agar: when agar is added (agar itself is not a nutrient)
Anaerobic Growth Media and Methods
>reducing media are used
>contains sodium thioglycollate
>ordinary, tightly capped test tubes
Special Culture Techniques
>Mycobacterium leprae (leprosy bacillus_ grown in armadillos for relatively low
body temp
>syphilis spirochete
>obligate intracellular bacteria, such as rickettsias and chlamydias, do not gro
w on artificial media. like viruses, they can reproduce only in a living host ce
ll
>special CO2 incubators for capnophiles, microbes that grow better at high CO2 c
onc.
Selective and Differential Media
>selective media: designed to suppress the growth of unwanted bacteria and encou
rage the growth of the desired microbes; example: bismuth sulfite agar used for
G(-) Salmonella typhi (typhoid fever) from feces
>differential media: make it easier to distinguish colonies of the desired organ
ism from other organism growing on the same plate; example: blood agar (contains
RBCs) is a medium used to identify bacterial species that destroy RBCs. Strepto
coccus pyogenes, cases strep throat, have clear ring around their colonies where
they have lysed the surrounding RBCs
Enrichment Media
>used bec. bacteria present in small numbers can be missed, esp. if other bacter
ia are present in much larger numbers
>usu. liquid & provides nutrients and environmental conditions that favor the gr
owth of a particular microbe but not others; also a selective medium
Obtaining a Pure Culture
<refer to ppt>
The Growth of Bacterial Cultures
<refer to ppt>
>when the number of cells in aech generation is expressed as a power of 2, the e
xponent tells the number of doublings (generations) that have occurred
>bacterial growth curve shows the growth of cells over time: lag, log, stationar
y, death
>because the generation time is constant, a logarithmic plot of growth during th
e log phase is a straight line
Direct Measurement of Microbial Growth
<refer to ppt>
>plate counts are often reported as colony-forming units (CFU)
>serial dilution
>pour plates and spread plates
>filtration - when the quantity of bacteria is very small, as in lakes or relati
vely pure streams; applied frequently to detection and enumeration of coliform b
acteria, which are indicators of fecal contamination of food or water
>Most Probable Number (MPN) Method: statistical estimating technique based on th

e fact that the greater the number of bacteria in a sample, the

needed to reduce the density to the point at which no bacteria
in the tubes in a dilution series; the MPN is only a statement
95% chance that the bacterial population falls within a certain
he MPN is statistically themost probable number

more dilution is
are left to grow
that there is a
range and that t

Estimating Bacterial Numbers by Indirect Methods

>Turbidity
>Metabolic Activity
>Dry Weight; for filamentous bacteria and molds
Chapter 8: Microbial Genetics
Structure and Function of the Genetic Material
>genetics: the science of heredity
>genome: genetic information in a cell; includes its chromosomes and plasmids
>chromosomes: structures containing DNA that physically carry hereditary informa
tion; they contain the genes
>genes: segments of DNA that code for functional products
>DNA: macromolecule composed of repeating units called nucleotides (each nucleot
ide = nucleobase + deoxyribose + phosphate grp)
>each strand has a string of alternating sugar and phosphate groups (its sugar-p
hosphate backbone), and a nitrogenous base is attached to each sugar
>the 2 strands are held together by H-bonds b/n their nitro bases
>base pairs: adenine + thymine; cytosine + guanine
>genetic code: set of rules that determines how a nucleotide sequence is convert
ed into the amino acid sequence of a protein
>2 primary features of biological information storage:
1. the linear sequence of bases provides the actual information
2. the complementary structure allows for the precise duplication of
DNA during cell dividion
>when the ultimate molecule for which a gene codes has been produced, we say tha
t the gene has been expressed
Genotype and Phenotype
>genotype: genetic makeup; the information that codes for all the particular cha
racteristics of the organism; represents potential properties
>phenotype: refers to actual expressed properties; manifestation of genotype
>in molecular terms: genotype = colleciton of genes, its entire DNA; phenotype =
collection of proteins
>in microbes, most proteins are either enzymatic (catalyze particular reactions)
or structural (participate in large functional complexes like membranes or flag
ella)
DNA and Chromosomes
>bacteria typically have a single circular chromosome consisting of a single cir
cular molecule of DNA with associated proteins
>chromosome is looped & folded & attached to the PM
>genomics: the sequencing and molecular characterization of genomes
DNA Replication
>bc of the bases are complementary, one strand of the double-helical DNA can act
as a template for the production of the other strand
---p. 47---Nucleic Acids
>1944: Avery, MacLeod, McCarty discovered that a substance called DNA is the sub
stance of which genes are made
>Watson & Crick (used data of Wilkins & Franklin) identified the physical struct
ure of DNA
>Crick suggested a mechanism for DNA replication

>nucleotides are the structural units of nucleic acids

>Each nucleotide has 3 parts: a N-containing base, a pentose (5-carbon) sugar, e
ither deoxyribose or ribose, a phosphate group (phosphoric acid)
>N-containing bases are cyclic compounds comp. of C, H, O, N atoms
>purines: G, A
>pyrimidines: C, U, T
>nucleoside: combination of a purine/pyrimidine plus a pentose sugar; no phospha
te group
<refer to figure>
DNA
>2 long strands wrapped around each other to form a double helix
>every strand has a backbone of alternating deoxyribose sugar & phosphate groups
; the deoxyribose of one nucleotide is joined to the phosphate group of the next
---p.213--->when replication beginds, the supercoiling is relaxed by topoisomerase or gyras
e, and the 2 strands of parental DNA are unwound by helicase
>free nucleotides present in the cytoplasm are matched up to the exposed bases o
f the single-stranded parental DNA
>any bases that are improperly base-paired are removed and replaced by replicati
on enzymes
>the newly added nucleotide is joined to thr growing DNA strand by DNA polymeras
e
>the parental DNA is unwound a bit further
>replication fork: the point at which replication occurs
>semiconservative replication: each new double-stranded DNA molcule contains one
original/conserved strand & one new strand
>the end with the hydroxyl attached to the 3'C is called the 3' end of the DNA s
trand
>the end having a phosphate attached to the 5'C is called the 5' end
>this structure of DNA affects the replication process bc DNA polymerases can ad
d new nucleotides to the 3' end only
>as the replication fork moves along, the 2 new strands must grow in different d
irections
>DNA replication needs energy -> supplied by the nucleotides, w/c are actually n
ucleoside triphosphates -> 2 phosphate grps are removed to add the nucleotide to
a growing strand of DNA; hydrolysis of the nucleoside is exergonic
>DNA replication by some bacteria like E. coli goes bidirectionally around the c
hromosome; 2 RFs move in opp directions away from the origin of replication; RFs
eventually meet bc the bacterial chromosome is a closed loop
>accuracy of replication is largely due to the proofreading capability of DNA po
lymerase
https://www.youtube.com/watch?v=2iVltkYy0jg
https://www.youtube.com/watch?v=27TxKoFU2Nw
DNA Gyrase Relaxes supercoiling ahead of the replication fork
DNA Ligase Makes covalent bonds to join DNA strands: joins Okazaki fragments an
d new segments in excision repair
DNA Polymerase Synthesizes DNA: proofreads and repairs DNA
Endonucleases Cut DNA backbone in a strand of DNA: facilitate repair and inserti
ons
Exonucleases Cut DNA from an exposed end of DNA: facilitate repair
Helicase Unwinds double-stranded DNA
Methylase Adds methyl group to selected bases in newly made DNA
Photolyase Uses visible light energy to separate UV-induced pyrimidine dimers
Primase Makes RNA primers from a DNA template
Ribozyme RNA enzyme that removes introns and splices exons together
RNA Polymerase Copies RNA from a DNA template
snRNP RNA-protein complex that removes introns and splices exons together
Topoisomerase Relaxes supercoiling ahead of the replication fork: separates DNA
circles at the end of DNA replication
Transposase Cuts DNA backbone leaving single-stranded "sticky ends"

RNA and Protein Synthesis

Transcription
>the synthesis of a complementary strand of RNA from a DNA template
>mRNA, rRNA, tRNA
>the proces of transcription required both an enzyme called RNA polymerase and a
supply of RNA nucleotides
>transcription begins when RNA polymerase binds to the DNA at a site called the
promoter
>like DNA, RNA is synthesized in the 5' -> 3' direction
>RNA synthesis continues until RNA polymerase reaches a site on the DNA called t
he terminator
Translation
>protein synthesis; involves decoding the "language" of nucleic acids and conver
ting that info to the "language" of proteins
>the language of mRNA is in the form of codons, grps of 3 nucleotides
>each codon codes for a particular amino acid; this is the genetic code
>degeneracy: AA are signaled by several alternative codons; allows for certain a
mt of change or mutation in the DNA w/out affecting the protein
>61 sense codons (code for AA); 3 nonsense codons (stop codons)--UAA, UAG, UGA
>AUG, methionine -> start
>in bacteria, the start AUG codes for formylmethionine rather than methionine fo
und in other parts of the protein
>the initiating methionine is often removed later, so not all proteins begin wit
h methionine
>site of translation is the ribosome; tRNA molecules recognize the codons & tran
sport the required AA
>each tRNA molecule has an anticodon, w/c it can base-pair with its associated c
odon; each tRNA can also carry on its other end the AA encoded by the codon that
the tRNA recognizes
>ribosomes direct the orderly binding of tRNAs to codons & assemble the AA into
chains
>ribosomes: E, P, A sites
>AA joined via peptide bonds
>the ribosome moves along the mRNA in the 5' -> 3' direction
>in prokaryotic cells, the translation of mRNA into protein can begin even befor
e transcription is complete; bc mRNA is produced in the cytoplasm, the start cod
ons of an mRNA being transcribed are available to ribosomes before the entire mR
NA molcule is even made
>in eukaryotic cells, the mRNA must be completely synthesized & moved through th
e nuclear membrane to the cytoplasm before translation can begin; it also underg
oes processing before it leaves the nucleus
>in eukaryotic cells, the regions of genes that code for proteins are often inte
rrupted by noncoding DNA
>exons: expressed; introns: intervening regions of DNA that do not encode protei
n
>in the nucleus, RNA polymerase synthesizes RNA transcript that contains copies
of the introns
>snRNPs (small nuclear ribonucleoproteins) remove the introns and splice the exo
ns together; in some, the introns act as ribozymes
The Regulation of Bacterial Gene Expression
>cells save energy by making only those proteins needed at a particular time
>many genes, around 60-80% are not regulated but are constitutive (products cons
tantly produced); example: enzymes of glycolysis
Repression and Induction
These 2 genetic control mechanisms regulate the transcriptionof mRNA and consequ
ently the synthesis of enzymes from them

Repression
>inhibits gene expression & decreases the synthesis of enzymes
>response to the overabundance of an end-product of a metabolic pathway
>causes a decrease in the rate of synthesis of the enzymes leading to that produ
ct
>regulatory proteins called repressors, block the ability of RNA polymerase to i
ntiate transcription
>default position of a repressible gene is ON
Induction
>turns on the transcription of a gene
>inducer: substance that acts to induce transcription of a gene
>inducible enzymes: enzymes that are synthesized in the presence of inducers
>default position of an inducible gene is OFF
>example: genes required for lactose metabolism in E. coli; if E. coli is placed
in a mediam with no lactose, the organism contain almost no B-galactosidase; wh
en lactose is added, it produces a large quantity of the enzyme; lactose in the
cell is converted to allolactose, the inducer of these genes
The Operon Model of Gene Expression
>describes the details of the control of gene expression by induction & repressi
on
>formulated by Francois Jacob and Jacques Monod in 1961; based on lactose catabo
lism in E. coli
>in addition to B-galactosidase (splits lactose into glucose & galactose), enzym
es include lac permease (transport of lactose into the cell), transacetylase (me
tabolizes certain disaccharides other than lactose)
>the genes for the 3 enzymes are next to each other on the bacterial chromosome
& are regulated together
>in E. coli: genes for the 3 enzymes are in the lac operon; B-galactosidase is e
ncoded by lacZ; lac permease by lacY; transacetylase by lacA (whose fxn in lacto
se metabolism is still unclear) -> inducible operon
<refer to figure on p. 224>
>these genes are called structural genes bc they determine the structures of pro
teins
>when lactose is introduced into the culture medium, the lac structural genes ar
e all transcribed and translated rapidly and simultaneously
>control region -> promoter, the region of DNA where RNA polymerase initiates tr
anscription; operator, acts as a go or stop signal for transcription of the stru
ctural genes
>operon: set of operator & promoter sites + structural genes they control
>regulatory gene or I gene encodes a repressor protein that switches inducible a
nd repressible operons on or off
>lac operon is an inducible operon; repressor binds to the operator site, preven
ting transcription; if lactose is present, it binds to allolactose (inducer), en
zymes are transcribed
<refer to figure on p. 225>
>in repressible operons, the structural genes are transcribed until they are tur
n off or repressed
>genes of the enzymes for trytophan synthesis of E. coli (E. coli trp operon)
>when excess trp is present, it acts as a corepressor binding to the repressor p
rotein
>repressor protein bind to operator, stopping trp synthesis
Positive Regulation
>regulation of lac operon depends on glucose levels -> controls intracellular le
vel of cAMP (derived from ATP and serves as a cellular alarm signal)
>enzymes that catabolize glucose are constitutive (preferred nutrient is glucose
)

>when glucose is no longer available, cAMP acculumates

>cAMP binds to catabolic activator protein (CAP)
>CAP binds to the lac promoter -> initiates transcription by making it easier fo
r RNA polymerase to bind to the promoter
>thus, transcription of lac operon requires BOTH the presence of lactose and abs
ence of glucose
>cAMP is an alarmone, a chemical alarm signal that promotes a cell's response to
envtal or nutritional stress
>catabolite repression: inhibition of the metabolism of alternative carbon sourc
es by glucose; aka glucose effect
Mutation: Change in the Genetic Material
>mutation: change in the base sequence of DNA
>many simple mutations are silent (neutral) -> one nucleotide is substituted for
another in the DNA (esp. the 3rd position of the codon) -> denegeracy of the ge
netic code
Types of Mutations
>base substitution: most common type of mutation involving single base pairs; ak
a point mutation; a single base at one point in the DNA is replaced with a diffe
rent base. when the DNA replicates, the result is a substituted base pair -> thi
s mutation leads to an altered protein in the granddaughter cell -> missense mut
ation (if the base substitution results in an AA substition in the synthesized p
rotein)
>sickle cell disease is caused by a single change in the gene for globin; missen
se mutation from A to T -> change from glutamic acid to valine in the protein
>nonsense mutation: base substitution resulting in a nonsense codon; effectively
prevents synthesis of a complete functional protein
>frameshift mutation: one or a few nucleotide pairs are deleted or inserted in t
he DNA
>Huntington's disease: progressive neurological disoerder caused by extra bases
inserted into a particular gene
>spontaneous mutations: occur spontaneously because of occasional mistakes durin
g DNA replication; occur in the absence of any mutation-causing agents
>mutagens: agents in the environment (like chemicals & radiation) that directly
or indirectly bring about mutations
>certain mutations result in resistance to antibiotics or altered pathogenicity
example: Salmonella enterica with an altered outer membrane can
survive in phagocytes
example: mutation in a capsule-encoding gene may result in decreased
pathogenicity bc phagocytes can destroy the bacteria, Streptococcus
pneumoniae, Haemophilus influenzae, Neisseria meningitidis
Mutagens
Chemical Mutagens
>nitrous acid (HNO2): converts the base adenine to a form that pairs with C inst
ead of T; like all mutagens, it alters DNA at random locations
>nucleoside analog: structurally similar to normal nitrogenous bases but have sl
ightly altered base-pairing properties; examples: 2-Aminopurine nucleoside in pl
ace of adenine (AT -> CG); 5-Bromouracil nucleoside in place of thymine (AT -> C
G); the analogs cause mistakes in base pairing -> subsequent replication has err
or
>benzopyrene, present in smoke & soot, is an effective frameshift mutagen
>aflatoxin, produced by Aspergillus flavus (mold on peanuts & grain) is a frames
hift mutagen
>frameshift mutagens are often potent carcinogens
Radiation
>X rays & gamma rays are potent mutagens bc of their ability to ionize atoms & m
olecules

>penetrating rays cause electrons to pop out of their shells -> can combine w/ b
ases in DNA (error in DNA replication & repair that produce mutations) -> or bre
akage of covalent bonds of the sugar-phosphate backbone, causes physical breaks
in chromosomes
>UV light: nonionizing component of ordinary sunlight (dec. by ozone layer) -> f
ormation of harmful covalent bonds between certain bases, e.g. adjacent thymines
form thymine dimers
>bacteria & other organisms have enzymes that can repair UV-induced damage:
photolyases: aka light-repair enzymes; use visible light energy
to separate dimer back to the original 2 Ts
nucleotide excision repair: not restricted to UV-induced damage; can
repair mutations from other causes (endonuclease -> exonuclease ->
DNA poly
merase -> DNA ligase)
>methylases: discovered by Hamilton Smith in 1970; answers how the incorrect bas
e could be distinguished from the correct base if it was not physically distorte
d like the thymine dimer; these enzymes add a methyl grp to selected bases soon
after a DNA strand is made
>UV light in humans -> thymine dimers in skin cells -> result in skin cancer
>xeroderma pigmentosum -> increased sensitivity to UV light; defect in nucleotid
e excision repair
The Frequency of Mutation
>mutation rate: probability that a gene will mutate when a cell divides; usually
stated as a power of 10 (usu. negative exponent bc mutation is rare)
>mutagen increases the spontaneous rate of mutation, w/c is about one in 10^6 re
plicated genes, by a factor of 10 to 1000 times (10^-5 to 10^-3)
Identifying Mutants
>mutants can be detected by selecting or testing for an altered phenotype
>experiments usually performed with bacteria
>positive (direct) selection: detection of mutant cells by rejection of the unmu
tated parent cells (penicillin-resistant bacteria)
>negative (indirect) selection: selects a cell that cannot perform a certain fun
ction, using the technique of replica plating (auxotrophic mutant cannot synthes
ize histidine)
>replica plating is an effective means of isolating mutants that require one or
more new growth factors
>auxotroph: any mutant microorganism having a nutritional requirement that is ab
sent in the parent
Identifying Chemical Carcinogens
>many known mutagens have been found to be carcinogens, substances that cause ca
ncer in animals, including humans
>Ames test: preliminary screening of potential carcinogens that uses bacteria as
carcinogen indicators; based on the observation that exposure of mutant bacteri
a to mutagenic substances may cause new mutations that reverse the effect (the c
hange in phenotype) of the original mutation. These are called reversions; speci
fically, the test measures the reversion of histidine auxotrophs of Salmonella
his(-) [mutants that have lost the ability to synthesize histidine] to his(+) ce
lls after treatment with a mutagen; used rat liver extract as a rich source of a
ctivation enzymes
Genetic Transfer and Recombination
>genetic recombination: exchange of genes b/n 2 DNA molecules to form new combin
ations of genes on a chromosome
>crossing over: process where a cell picks up a foreign DNA (donor DNA) and inse
rt into its chromosome; donor DNA has a nick -> aligns with complementary base p
airs in the recipient chromosome -> RecA protein catalyzes the joining of 2 stra
nds -> recipient chromosome contains new DNA -> strands resolved by DNA polymera
se and ligase -> donor DNA will be destroyed

>in eukaryotes, crossing over egenrally takes place during the formation of repr
oductive cells
>in bacteria, genetic recombinatin can happen in a number of ways
>in present-day microbes, recombiantion is more likely than mutation to be benef
icial
>vertical gene transfer: occurs when genes are passed from an organism to its of
fspring; like in plants & animals
>horizontal gene transfer: bacteria can pass their genes not only to their offsp
ring, but also laterally, to other microbes of the same generation; donor cell g
ives portion of its total DNA to a recipient cell & then it will be degraded by
cellular enzymes -> recipient cell will then be called a recombinant
>not a frequent event
Transformation in Bacteria
>transformation: genes are transferred from one bacterium to another as "naked"
DNA in solution
>first demonstrated by Frederick Griffith in 1928
>he worked on 2 strains of Streptococcus pneumoniae; one was a virulent
or pathogenic strain and has a polysaccharide capsule to
prevent
phagocytosis; the avirulent strain lacks the capsule
>subsequent investigations based on his research revealed that bacterial
transformation could be carried out without mice
>experiments by Avery, MacLeod, McCarty -> the component responsible for
transforming harmless S. pneumoniae into virulent strains was DNA; their result
s provided one of the conclusive indications that DNA was indeed the carrier of
genetic info
>in nature, some bacteria, perhaps after death & cell lysis, release their DNA i
nto the environment -> some bacteria encounter them -> recombinant cell
>transformation occurs naturally among very few genera of bacteria, including Ba
cillus, Haemophilus, Neisseria, Acinetobacter, and certain strains of Streptococ
cus and Staphylococcus
>transformation works best when the donot & recipient cells are very closely rel
ated
>competence: results from alterations in the cell wall that make it permeable to
large DNA molecules; when a recipient cell can take up the donor DNA, it is sai
d to be competent
>E. coli is NOT naturally competent for transformation -> needs lab treatments
Conjugation in Bacteria
>conjugation: mediated by one kind of plasmid
>plasmid: a circular piece of DNA that replicates independently from the cell's
chromosomes; but the plasmid's genes are usually not essential for cell growth
>plasmids responsible for conjugation are transmissible b/n cells during conjuga
tion
>differs from transformation in 2 ways:
1. conjugation required direct cell-to-cell contact
2. conjugating cells must generally be of opposite mating type; donor cells must
carry the plasmid, and recipient cell usually do not
>in G(-) bacteria, the plasmid carries genes that code for the synthesis
of sex pili
>in G(+) bacteria, they produce sticky surface molecules for direct
contact
>the plasmid is replicated during the transfer of a single-stranded copy of the
plasmid DNA to the recipient, where the complementary strand is synthesized
>E. coli conjugation: the F factor (fertility factor) was the first plasmid obse
rved to be transferred between cells during conjugation
a. When an F factor (a plasmid) is translerred from a donor (F+) to a re
cipient (F-), the F- cell is converted to an F+ cell.
b. When an F factor becomes integrated into the chromosome of an F+ cell
, it makes the cell a high frequency of recombination (Hfr) cell.

c. When an Hfr donor passes a portion of its chromosome into an F-recipi

ent, a recombinant F- cell results.
<refer to figure on p. 238>
Transduction in Bacteria
>transduction: bacterial DNA is transferred from a donor cell to a recipient cel
l inside a virus that infects bacteria, called a bacteriophage, or phage
>generalized transduction
<refer to figure on p. 239>
>specialized transduction: only certain bacterial genes are transferred
Plasmids and Transposons
>genetic elements that provide additional mechanisms for genetic change
>occur in both prokaryotic & eukaryotic organisms
Plasmids
>found mainly in bacteria but also in some eukaryotic microorganisms like Saccha
romyces cerevisiae
>The F factor is a conjugative plasmid that carries genes for sex pili and for
the transfer of the plasmid to another cell
>sometimes plasmids can be crucial to the survival & growth of the cell
>dissimilation plasmids: code for enzymes that trigger the catabolism of certain
unusual sugars and hydrocarbons
>other plasmids code for proteins that enhance the pathogenicity of a bacterium;
example: strain of E. coli that causes diarrhea -> carries plasmids for toxin p
roduction & attachment to intestinal cells
>other plasmids contain genes for the synthesis of bacteriocins, toxic proteins
that kill other bacteria
>Resistance factors (R factors) are plasmids that have significant medical impor
tance; carry genes that confer upon their host cell resistance
to antibiotics, heavy metals, or cellular toxins. Many R factors contain two gro
ups of genes:
1. resistance transfer factor (RTF) - includes genes for plasmid replication and
conjugation
2. r-determinant - has the resistance genes; codes for the production of enzymes
that inactivate certain drugs or toxic substances
>unlike eukaryotes, bacterial species can conjugate and transfer plasmids to oth
er species
Neisseria may have acquired its penicillinase-producing plasmid from
Streptococcus; Agrobacterium can transfer plasmids to plant cells
Transposons
>small segments of DNA that can move (be "transposed") from one region of a DNA
molecule to another
>discovered by Barbara McClintock in corn
>may move from one site to another site on the same chromosome or to another chr
omosome or plasmid
>fortunately, transposition occurs relatively rarely
>all transposons contain the information for their own transposition
>insertion sequences (IS): simplest transposons; contain only a gene that codes
for an enzyme (transposase, which catalyzes the cutting & resealing of DNA that
occurs in transposition) & recognition sites
>recognition sites: short inverted repeat sequences of DNA that the enzyme recog
nizes as recombination sites b/n the transposon & the chromosome
>complex transposons carry other genes not connected with the transposition proc
ess
>transposons provide a natural mechanism for the movement of genes from one chro
mosome to another
>because they may be carried between cells on plasmids or viruses, they can als
o spread from one organism-or even species-to another.

--MICROBIAL CONTROL
Clostridium tetani (endospores) - causative agent of tetanus
Fungal spores
Parasite cysts
Mycobacterium tuberculosis - causative agent of tuberculosis
Naked virus
Hepatitis B
Polio
Most vegetative bacteria
Parasite trophozoites
Enveloped viruses (Influenza A)
Fungi
G(-) bacteria > G(+) bacteria
Naked viruses > Enveloped viruses