2.

7 DNA Replication,
Transcription, and
Translation
IB Biology Year 2
Fall 2015
Thanks to Ms. Stuckey, modified by V. Azuree 2015

animation link on
picture

DNA replication

Semi-Conservative Replication
2.7 U1 The replication of DNA is semi-conservative and depends on complementary base pairing.

Post replication, DNA is semi-conservative

1 strand old
1 strand new
Original used as a template for the 2 new strands
Complementary base pairing
A-T
C-G

Evidence for Semi-Conservative Replication

2.7 NOS1 Obtaining evidence for scientific theories: Meselson and Stahl obtained evidence for the
semi-conservative replication of DNA.

Even if it seems right, we need to back up theories!

1958 - Meselson and Stahl
15
N DNA vs. 14N DNA
Caesium chloride and a centrifuge
Multiple generations of E. coli in 15N then transfer to
14
N
Measured density after several generations.
HUH?!?

Link to video in picture!

See also:
Bozeman Bio

Meselson and Stahl

Pause to Analyze
2.7 S2 Analysis of Meselson and Stahls results to obtain support for the theory of semi-conservative
replication of DNA.

p.113 -114 in your book

(see answer key if we dont get to it in class!)

Helicase (everyones fave enzyme)

2.7 U2 Helicase unwinds the double helix and separates the two strands by breaking hydrogen bonds.

Step #1
Twisty double-helix in your way? Here comes helicase!
Enzyme that uses ATP to unwind DNA by breaking Hbonds
Actually a protein complex
Looks like a donut (yum)
Untwist AND unzip

DNA REPLICATION !

DNA polymerase
2.7 U3 DNA polymerase links nucleotides together to form a new strand, using the pre-existing strand
as a template

Step #2
Moves from 5 3 - NO EXCEPTIONS
Leading strand vs. lagging strand
DNA polymerase adds in complementary nucleotides by
positioning them properly to form H bonds with the
template
Covalently bonds phosphate group to sugar.
Pretty fool-proof
https://www.youtube.com/watch?v=8kK2zwjRV0M#t=535

PCR - Polymerase Chain Reaction HUMAN MADE

2.7 A1 Use of Taq DNA polymerase to produce multiple copies of DNA rapidly by the polymerase chain reaction
(PCR).

Make MANY copies of DNA using change in temp.

High temp (~95C) breaks H bonds
Cool down (~54C) for re-annealing of primers
Extra primers prevent parents from coming back
together
Temp to 72C to allow Taq DNA polymerase to
replicate DNA
Taq DNA polymerase
withstand 95C, still works at 54C, optimum
72C
Repeat over and over
30 cycles - DNA amplified by 1 billion!!

PCR - Polymerase Chain Reaction

2.7 A1 Use of Taq DNA polymerase to produce multiple copies of DNA rapidly by the polymerase
chain reaction (PCR).

animation
linked to
picture

animation link on
picture

Transcription

Transcription
2.7 U4 Transcription is the synthesis of mRNA copied from the DNA base sequences by RNA
polymerase.

DNA is pretty much useless without being converted to

readable polypeptides.
First step between DNA Protein = Transcription

Transcription
2.7 U4 Transcription is the synthesis of mRNA copied from the DNA base sequences by RNA
polymerase.

Transcription
DNA (template) mRNA (end result)
occurs in the NUCLEUS
1. RNA polymerase binds at the start of a gene
2. Similar to DNA replication - RNA polymerase forms Hbonds between complementary base pairs and covalent
bonds between nucleotides
a. Still moves 5 3
3. RNA separates and double helix reforms
4. Transcription stops at the end of the gene.

Transcription
2.7 U4 Transcription is the synthesis of mRNA copied from the DNA base sequences by RNA
polymerase.

Sense vs. Antisense strands

Does it matter which strand DNA copies?
YES!!!
Template strand = ANTISENSE strand
is transcribed from 5 3
has the complementary DNA
sequence
Sense strand - not transcribed
Sequence matches mRNA (but T for
U!)

animation link on
picture

Translation

Translation
2.7 U5 Translation is the synthesis of polypeptides on ribosomes.

Step #2
mRNA polypeptide chain
Ribosome reads and translates mRNA codons into amino acid
chains
2 subunits - large and small
composed of rRNA, proteins

mRNA and the Genetic Code

2.7 U6 The amino acid sequence of polypeptides is determined by mRNA according
to the genetic code.

mRNA - all different lengths (av. 2,000 nucleotides)

depends on the gene
How much mRNA and which mRNA is produced?
depends on the polypeptides needed by the cell
Pancreas
lots of mRNA that carries code for insulin
Dont forget about tRNA and rRNA, too!

Codons (or, Yes, Jessica, you have to use the stupid chart thingie)
2.7 U7 Codons of three bases on mRNA correspond to one amino acid in a polypeptide.

How does the tRNA know which amino

acids to bring?
Codons!
mRNA read in sets of 3 nucleotides
that code for a specific amino acid.
64 possibilities (4x4x4)
20 amino acids
Memorize them all?
Heck no! CODON CHART!

Decoding Base Sequences

2.7 S1, S3, S4 Use a table of the genetic code to deduce which codon(s) correspond to which amino acids; Use a table of
mRNA codons and their corresponding amino acids to deduce the sequence of amino acids coded by a short mRNA
strand of known base sequence; Deducing the DNA base sequence for the mRNA strand.

1. Deduce the codons for:

a. Trp UGG
b. Tyr UAU, UAC
c. Arg
CGU, CGC, CGA, AGA, AGG
2. Deduce amino acid sequence that corresponds to these mRNA sequences:
a. ACG Thr
b. CACGGG His Gly
c. CGCGCGAGG Arg Ala Arg
3. If the mRNA contains the base sequence CUC AUC GAA UAA CCC
a. deduce the AA sequence of the polypeptide translated from the mRNA
Leu sequence
Ile Glu STOP
b. deduce the base
ofPro
the ANTISENSE strand of DNA
GAG TAG CTT ATT GGG

Codons and Anticodons

2.7 U8 Translation depends on complementary base pairing between codons on mRNA and
anticodons on tRNA.

1. mRNA has a 3 nucleotide

codon
2. tRNA has a 3 nucleotide
complementary anticodon
3. Ribosomes are binding sites
for both and catalyse
polypeptide assembly

Codons and Anticodons

2.7 U8 Translation depends on complementary base pairing between codons on mRNA and
anticodons on tRNA.

Main events:
1. mRNA binds to small subunit of ribosome
2. tRNA brings the first AA
3. 2nd tRNA brings the next AA. Only 2 tRNAs at a time!
4. Ribosome catalyzes the addition of the 2 AAs
5. Ribosome slides down, releasing tRNA 1 and shifting tRNA 2
6. Next tRNA comes in
7. Ribsome makes next peptide bond
Repeat over and over!
Mistakes are rare!

Production of Human Insulin in Bacteria

2.7 A2 Production of human insulin in bacteria as an example of the universality of the genetic code
allowing gene transfer between species.

Diabetes patients need insulin

bovine, porcine, shark used in past
has some differences!
can cause allergies
Genetically transfer the human insulin gene into bacteria
Bacteria then mass produce the human version of
insulin which we can harvest!

More review:
Learn Genetics Transcribe and Translate a Gene
DNA, Hot Pockets, & The Longest Word Ever: Crash Course Biology #11
Bozeman Science DNA Replication
Bozeman Science Transcription and Translation