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TUTORIAL: DNA BIOLOGY AND TECHNOLOGY 1.Describe the biochemical composition, str
ucture and replication of DNA. DNA is a polymer where the monomer units of DNA a
re nucleotides and the
polymer is known as polynucleotide.DNA molecule consists of two strands of
polynucleotide that wound together into a double helix.In the two polynucleotide
strands
is a phosphate group linked together by a phosphodiester bond between the phosph
ate
group of one nucleotide and the hydroxyl group of carbon 3 of the next nucleotid
e that is
3 terminal while at one end of each strand is a phosphate group linked to the car
bon 5 of
deoxyribose sugar.This is the 5 terminal of each strand.The 3 and 5 terminals of the
two strands are at opposite ends where the two strands is antiparallel.The two
polynucleotide strands in the double helix DNA molecule are held together by hyd
rogen
bonds between complementary purine-pyrimidine base pairs.There are four differen
t
types of nucleotides found in DNA, differing only in the nitrogenous which are a
denine,
guanine, cytosine and thymine.
The sequence of steps in DNA replication is based on the semiconservative model
that is takes place in the 5 -3 direction using a DNA template strand.This process
is
catalyzed by DNA polymerase III.This process begin with the unwinding of the DNA
double helix to form a replication fork.The unwinding is catalyzed by the enzyme
, DNA
helicase.The DNA helicase enzyme unwinds the double-stranded DNA to a replicatio
n
fork ready for replication.The antiparallel nature of the strands, that is the 5 -
3
orientation of the top strand and the 3 -5 orientation of the complementary bottom
strand.Then, the DNA is already partially unwound to form a replication fork.On
the
bottom template strand, RNA primase attaches a short RNA primer in the 5 -3 directi
on.
After that, the RNA primase is removed, and free DNA nucleosides are added by DN
A
poltmerase III to the RM|NA primer in the 5 -3 direction.This new strand is called
the
leading strand because it is made in the same direction as the movement of the r
eplication
fork.On the top template strand, RNA primase synthesizes a short RNA primer in t
he a
short RNA primer in the 5 -3 direction.RNA primase is removed, and DNA polymerase
III adds nucleosides to the RNA primer one after another opposite to the replica
tion fork
to form a short length DNA.This new short length is called the lagging strand be
cause it
is made in the direction opposite to the movement of the replication fork.The fr
agment
produced is also called Okazaki fragment.The process repeat as the DNA continues
to
unwind. Then, the RNA primers are removed by a different type of enzyme called D
NA
polymerase I.The two Okazaki fragments are then sealed and joined up by the DNA
ligase to produce a continuous chain called lagging strand antiparallel to the l
eading
strand formed earlier.Two new antiparallel continuous strands which are one lead
ing
strand and one lagging strand, are formed.The new double-stranded daughter DNA
molecules daughter DNA molecules then twist or wind to form double helix DNA
molecules.
2.(a)Describe the steps in the synthesis of the polypeptide. There are two proce
sses that undergoes by a cell in the synthesis of the polypeptide which are tran
scription and translation.Transcription is a process where the information in DN
A is transcribed to a RNA molecule, called messenger RNA (mRNA). During transcri
ption, one of the DNA strands acts as a template whereby a mRNA is transcribed c
omplementary to the DNA template strand.A specific protein-coding gene consists
of a promoter followed by the RNA-coding sequences for a protein and then a term
inator.The promoter is a base-sequence that specifies where transcription is beg
ins while the RNA-coding sequences is a base-pair sequence that includes coding
information for the polypeptide chain specified by the gene.Besides, the termina
tor is a sequence that specifies the end of the mRNA transcript.Transcription is
catalyzed by an enzyme called RNA polymerase when initiation of the transcripti
on at the promoter site is begins.This involves specific recognition of promoter
base sequence by RNA polymerase (in prokaryotes) or a complex of proteins (in e
ukaryotes).RNA synthesized is initiated with the addition of free RNA nucleotide
s in the 5 -3 direction opposite with attachment at -OH end to the uncoiling direct
ion of the DNA segment.The same rule of complementary base pairing are followed
as in replication, except that uracil (U) replaces thymine (T) to pair with aden
ine (A), as RNA does not contain tnymine.After the RNA
synthesis is initiated, the RNA elongation will continue to the direction of 5 -3 d
irection with the addition of more free nucleotides by RNA polymerase.Then, tran
scription will terminate when the terminator base sequence is recognized by RNA
polymerase.In eukaryotes, introns and exons formed in the pre-mRNA during transc
ription are removed to produce mature mRNAs. While translation is a process wher
e the genetic information transferred to a mRNA from the DNA is translated by ri
bosomes into amino acid sequence on polypeptides.This process requires the assis
tance of tRNA including the mRNA, rRNA and ribosome, enzymes and ATP which are i
nitiated with the movement of mRNA from nucleus to ribosome.Ribosomes, the organ
elles on which the mRNA is translated, consist of two subunits, each of which co
ntains rRNA an ribosomal protein.Ribosomes do not carry genetic information but
facilitate the interaction of tRNA with mRNA during protein synthesis.The small
subunit of a ribosome, binds to the 5 end of an mRNA molecule.The small subunit s
lides along the mRNA until it reaches the start codon, which shows where transla
tion should be initiated.The large subunit then binds to the small subunit to fo
rm a complete RNA-ribosome complex.The first tRNA occupies the P site and the A
site is available for the next tRNA.At the end of the initiation stage, elongati
on begins when the vacant A site is bound by another charged tRNA with an antico
don complementary to the codon after the start codon on the mRNA.The ribosome mo
ves three (a codon) nucleotides along the mRNA in a 5-3 direction and detaches the
peptide from the tRNA at the P site.The detached peptide is then attached by a
peptide linkage to the single amino acid on the A site.In the process during whi
ch the ribosome slides across to the next codon of the mRNA, the tRNA at the P s
ite is then displaced and detached from the ribosome.The P site is then adjacent
tRNA carrying the polypeptide chain.The vacant A site will the be occupied by t
he next tRNA.The steps repeat in a cycle called ribosome until translation is te
rminated.When the ribosome reaches a stop codon, there is no tRNA that has a com
plementary anticodon.This signals the end of translation.The large subunit advan
ces over the small subunit.The polypeptide is detached from the tRNA, and starts
to fold up to form the final shape of the protein.The rRNA disintegrates to rel
ease the large subunit, small subunit, tRNA and mRNA.
TUTORIAL: DNA BIOLOGY and TECHNOLOGY 1. Describe the Biochemical Composition,
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