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![Page 1: From Gene to Protein Chapter 17. What Is a Gene? The idea of the gene has evolved through the history of genetics We have considered a gene as – A discrete.](https://reader035.fdocuments.net/reader035/viewer/2022081603/56649e675503460f94b626b1/html5/thumbnails/1.jpg)
From Gene to Protein
Chapter 17
![Page 2: From Gene to Protein Chapter 17. What Is a Gene? The idea of the gene has evolved through the history of genetics We have considered a gene as – A discrete.](https://reader035.fdocuments.net/reader035/viewer/2022081603/56649e675503460f94b626b1/html5/thumbnails/2.jpg)
What Is a Gene? • The idea of the gene has evolved through the
history of genetics• We have considered a gene as
– A discrete unit of inheritance – A region of specific nucleotide sequence in a
chromosome– A DNA sequence that codes for a specific
polypeptide chain
![Page 3: From Gene to Protein Chapter 17. What Is a Gene? The idea of the gene has evolved through the history of genetics We have considered a gene as – A discrete.](https://reader035.fdocuments.net/reader035/viewer/2022081603/56649e675503460f94b626b1/html5/thumbnails/3.jpg)
Proteins= link between genotype and phenotype
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Flow of Genetic Information
• DNA= “directions” are encoded in nucleotide sequences– Sequences provide information for protein
synthesis• Gene expression= process by which DNA
directs protein synthesis– 2 stages: transcription and translation
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Flow of Genetic Information• Central Dogma
– Concept that cells are governed by a cellular chain of command: DNA RNA protein
• RNA= link between genes and the proteins• Transcription= synthesis of RNA under the
direction of DNA– messenger RNA (mRNA)
• Translation= synthesis of a polypeptide, using information in the mRNA
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Flow of Genetic Information
DNA
RNA
Protein
![Page 7: From Gene to Protein Chapter 17. What Is a Gene? The idea of the gene has evolved through the history of genetics We have considered a gene as – A discrete.](https://reader035.fdocuments.net/reader035/viewer/2022081603/56649e675503460f94b626b1/html5/thumbnails/7.jpg)
DNA
mRNARibosome
Polypeptide
TRANSCRIPTION
TRANSLATION
TRANSCRIPTION
TRANSLATION
Polypeptide
Ribosome
DNA
mRNA
Pre-mRNARNA PROCESSING
(a) Bacterial cell (b) Eukaryotic cell
Nuclearenvelope
• Bacteria– Translation can begin before
transcription finishes mRNA molecule
• Eukaryotes– Transcription occurs in nucleus– Translation occurs in cytoplasm
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Variation in Transcription and Translation
• Bacteria and eukaryotes differ in..– RNA polymerases– Termination of transcription– Ribosomes
• Archaea are prokaryotes, but share many features of gene expression with eukaryotes
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Form = Function• High diversity of molecules due to arrangement
of monomers– DNA composed of 4 nucleotides – RNA composed of 4 nucleotides – 20 amino acids (monomers of proteins)
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Form = Function• The flow of information from gene to protein is
based on a triplet code – Nonoverlapping, three-nucleotide codon
• Codons of a gene are…– Transcribed into complementary codons of mRNA– Translated into amino acids
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Second mRNA base
Firs
t mRN
A ba
se (5
end
of c
odon
)
Third
mRN
A ba
se (3
end
of c
odon
)
UUU
UUC
UUA
CUU
CUC
CUA
CUG
Phe
Leu
Leu
Ile
UCU
UCC
UCA
UCG
Ser
CCU
CCC
CCA
CCG
UAU
UACTyr
Pro
Thr
UAA Stop
UAG Stop
UGA Stop
UGU
UGCCys
UGG Trp
GC
U
U
C
A
U
U
C
C
CA
U
A
A
A
G
G
His
Gln
Asn
Lys
Asp
CAU CGU
CAC
CAA
CAG
CGC
CGA
CGG
G
AUU
AUC
AUA
ACU
ACC
ACA
AAU
AAC
AAA
AGU
AGC
AGA
Arg
Ser
Arg
Gly
ACGAUG AAG AGG
GUU
GUC
GUA
GUG
GCU
GCC
GCA
GCG
GAU
GAC
GAA
GAG
Val Ala
GGU
GGC
GGA
GGGGlu
Gly
G
U
C
A
Met orstart
UUG
G
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Cracking the Code• All 64 codons were deciphered by the mid-
1960s• Of the 64 triplets
– 61 codons represent amino acids– 3 codons are “stop” signals
• Genetic code– Redundant
• 1 amino acid coded by multiple codons
– Not Ambiguous• 1 codon = 1 amino acid
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Cracking the Code
• Codons must be read in the correct reading frame– 5’ to 3’ direction
• 5’-AUGCUGGAACCGACCUGA-3’
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Evolution of the Genetic CodeGenetic code is nearly universal…
From unicellular organisms….
..to multicellular animals
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Evolution of the Genetic CodeGenes can be transcribed and translated after being transplanted from one species to another
Tobacco plant expressing firefly gene
Pig expressing jellyfish gene
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Green sea slug expresses algal DNA• Green sea slugs (Elysia chlorotica) have functional
chloroplasts that carry out photosynthesis– Chloroplasts are taken up from food source= algae– Algae DNA incorporated into slug DNA
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Transcription
• Occurs in Nucleus• RNA made from DNA directions
– Messenger RNA (mRNA)• Only one strand of DNA transcribed
– Template strand= 3’ to 5’– RNA made in 5’ to 3’ direction
DNA
RNA
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Transcription• Enzyme= RNA polymerase• Purpose of RNA polymerase
– Separates DNA strands– Links RNA nucleotides together– Several dozen nucleotide pairs in length
• Section of DNA transcribed = transcription unit
DNA
RNA
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Transcription
• mRNA= complementary to DNA template strand– same base-pairing rules as DNA,
with one exception• Uracil substitutes for thymine• Uracil-Adenine
• Eukaryotes– Primary transcript= Initial RNA molecule produced
by transcription• Additional processing needed to make final mRNA
DNA
RNA
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Transcription: Initiation
• Promoter= nucleotide sequence signals binding site for RNA polymerase
• Transcription initiation complex= RNA polymerase II + transcription factors – Transcription factors= proteins control translation
of a gene by activating or preventing binding of RNA polymerase to promoter site
• A promoter called a TATA box is crucial in forming the initiation complex in eukaryotes
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Figure 17.8
Transcription initiationcomplex forms
3
DNAPromoter
Nontemplate strand
53
53
53
Transcriptionfactors
RNA polymerase IITranscription factors
53
53
53
RNA transcript
Transcription initiation complex
5 3
TATA box
T
T T T T T
A A A A A
A A
T
Several transcriptionfactors bind to DNA
2
A eukaryotic promoter1
Start point Template strand
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Transcription: Elongation• RNA polymerase untwists the double helix
– 10-20 bases at a time– Eukaryotes Rate of Transcription=
40 nucleotides/second• Nucleotides= added to the 3 end of the
growing RNA molecule
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Nontemplatestrand of DNA
RNA nucleotides
RNApolymerase
Templatestrand of DNA
3
35
5
5
3
Newly madeRNA
Direction of transcription
A
A A A
AA
A
T
TT
T
TTT G
GG
C
C C
CC
G
C CC A AA
U
U
U
end
Figure 17.9
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Transcription: Termination • Prokaryotes
– Terminator= nucleotide sequence signaling end of gene, RNA polymerase separates from DNA
– mRNA does not need additional modification– Translation can begin immediately
• Eukaryotes– Polyadenylation signal sequence= RNA transcript
released 10–35 nucleotides past this sequence– Additional processing needed to convert transcript
into functional mRNA molecule– Processing occurs before leaves nucleus
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Figure 17.7-4 Promoter
RNA polymeraseStart point
DNA
53
Transcription unit
35
Elongation
53
35
Nontemplate strand of DNA
Template strand of DNARNAtranscriptUnwound
DNA2
3535
3
RewoundDNA
RNAtranscript
5
Termination3
35
5Completed RNA transcript
Direction of transcription (“downstream”)
53
3
Initiation1
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RNA Processing
• Enzymes in the eukaryotic nucleus modify RNA transcript before released to cytoplasm– Alteration of ends of the primary transcript – RNA splicing of interior portions of molecule
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RNA Processing• Each end of a pre-mRNA molecule is modified
in a particular way– 5 end= modified nucleotide 5 cap– 3 end= poly-A tail
• Reasons for modifications– Facilitate export of mRNA from nucleus– Protect mRNA from enzymes in cytoplasm– Assist with ribosomal attachment to 5 end
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RNA Processing• Most eukaryotic genes have introns between
coding regions– Introns= series of nucleotides in noncoding regions
of genes – Included in RNA transcripts
• Exons= coding regions of genes – Expressed when translated into amino acid
sequences• RNA splicing removes introns and joins exons
– Spliceosomes– Ribozymes
• End product= mRNA molecule with continuous coding sequence
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What is the purpose of introns?
• Sequences may regulate gene expression• Genes can encode more than one kind of
polypeptide– Type of polypeptide depends on which segments are
removed during splicing– Alternative RNA splicing
• Advantage= Number of different proteins produced is much greater than its number of genes
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RNA ProcessingSpliceosomes= proteins + small nuclear ribonucleoproteins (snRNPs)
– Recognize splice sites
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RNA Processing• Ribozymes= catalytic RNA molecules
– Function as enzymes – Splice RNA– Discovery changed long-held belief that all biological
catalysts were proteins• Three properties of RNA enable it to function as an
enzyme– Form a 3-D structure
• Base-pair with itself
– Bases contain functional groups that act as a catalyst– Hydrogen-bond with other nucleic acid molecules
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5 Exon Intron Exon5 CapPre-mRNA
Codonnumbers
130 31104
mRNA 5Cap
5
Intron Exon
3 UTR
Introns cut out andexons spliced together
3
105 146
Poly-A tail
Codingsegment
Poly-A tail
UTR1146
RNA Processing
Final Product
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Flow of Genetic Information
Transcription
DNA
RNA
Protein
Translation
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Transcription Translation• Flow of information
from DNA to protein is the triplet code of bases
• mRNA moves out of nucleus and into cytoplasm
• Ribosomes attach to mRNA and translation begins
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Translation•Ribosomes “read” code on mRNA to build polypeptide chain •Amino acids brought to ribosome by transfer RNA (tRNA)– Single RNA strand – ~80 nucleotides long
RNA
Protein
Ribosomes
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TranslationMolecules of tRNA pair with a specific amino acid
• 3’ End= Amino acid attachment site
• Anticodon– Base-pairs with
complementary codon on mRNA
• Hydrogen bonds give tRNA its 3-D structure– L-shaped
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Translation
• Accurate translation requires 2 steps:1. Match between tRNA and an amino acid
– Enzyme aminoacyl-tRNA synthetase
2. Match between tRNA anticodon and mRNA codon
• Wobble= flexible pairing at the third base of a codon – Allows some tRNAs to bind to more than one
codon
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Aminoacyl-tRNAsynthetase (enzyme)
Amino acid
P P P Adenosine
ATP
P
P
P
PPi
i
i
Adenosine
tRNA
AdenosineP
tRNA
AMP
Computer model
Aminoacid
Aminoacyl-tRNAsynthetase
Aminoacyl tRNA(“charged tRNA”)
Figure 17.16-4
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Ribosomes• Facilitate specific coupling of tRNA anticodons
with mRNA codons– Two ribosomal subunits- large and small
• Proteins and ribosomal RNA (rRNA)
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Ribosomes
• Two types of ribosomes – Free ribosomes= in the cytosol– Bound ribosomes= attached to the endoplasmic
reticulum (ER)– Both types are identical – Switch from free to bound
• Free ribosomes mostly synthesize proteins that function in the cytosol
• Bound ribosomes make proteins of the endomembrane system and proteins to be secreted from the cell
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Ribosomes
• Polypeptide synthesis always begins in the cytosol
• Finishes in the cytosol unless the polypeptide signals the ribosome to attach to the ER
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Ribosomes
• Three binding sites for tRNA– P site= holds the tRNA with the growing
polypeptide chain– A site= holds the tRNA with next amino acid to
be added– E site= exit site
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Figure 17.17c
Amino end
mRNA
E
(c) Schematic model with mRNA and tRNA
5 Codons
3
tRNA
Growing polypeptide
Next aminoacid to beadded topolypeptidechain
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Translation: InitiationBrings together translation initiation complex:
– mRNA– tRNA with first amino acid– two ribosomal subunits
1. Small ribosomal subunit binds with mRNA and initial tRNA
2. Small subunit moves along the mRNA to start codon (AUG)
3. Proteins called initiation factors bring in the large subunit
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Translation: Initiation
• First tRNA attaches at P site• All other tRNA enter at A
site
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Translation: Elongation• Amino acids added one by one to the growing
chain– Proteins called elongation factors
• Occurs in three steps– Codon recognition– Peptide bond formation– Translocation
• Translation proceeds along the mRNA in a 5 to ′3 direction′
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Amino end ofpolypeptide
mRNA
5
E
Asite
3
E
GTP
GDP P i
P A
E
P A
GTP
GDP P i
P A
E
Ribosome ready fornext aminoacyl tRNA
Psite
Figure 17.19-4
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Translation: Termination
• Stop codon in the mRNA reaches the A site of the ribosome
• A site accepts a protein called a release factor• Release factor causes the addition of a water
molecule instead of amino acid– Reaction releases polypeptide– Translation assembly separates
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Translation• Polyribosome: multiple ribosomes translate a single
mRNA simultaneously– Enable cell to make many copies of a polypeptide very
quickly
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Completedpolypeptide
Incomingribosomalsubunits
Start ofmRNA(5 end)
End ofmRNA(3 end)(a)
Polyribosome
Ribosomes
mRNA
(b)0.1 m
Growingpolypeptides
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DNAtemplatestrand
TRANSCRIPTION
mRNA
TRANSLATION
Protein
Amino acid
Codon
Trp Phe Gly
5
5
Ser
U U U U U3
3
53
G
G
G G C C
T
C
A
A
AAAAA
T T T T
T
G
G G G
C C C G GDNAmolecule
Gene 1
Gene 2
Gene 3
C C
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Modifications of Proteins• During and after synthesis, a polypeptide
chain spontaneously coils and folds into its three-dimensional shape
• After translation, many proteins must undergo modifications before becoming functional– Activated by enzymes that cleave them– Multiple polypeptide chains come together to
form a larger protein
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Modifications of Proteins
• Targeted to specific sites in cell• Polypeptides destined for the ER or for secretion are
marked by a signal peptide
• A signal-recognition particle (SRP) binds to the signal peptide– Brings the signal peptide and its ribosome to the
ER– Signal peptide removed and protein enters ER for
transport
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Effect of Mutations on Proteins
• Mutations = changes in the genetic material of a cell– Can occur during DNA replication, recombination, or
repair– Mutagens= physical or chemical agents that can
cause mutations• Point mutations= changes in one base pair of a
gene– Single change in a DNA template strand can lead to
the production of an abnormal protein
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Wild-type hemoglobin
Wild-type hemoglobin DNA3
3
35
5 3
35
5553
mRNA
A AGC T T
A AGmRNA
Normal hemoglobin
Glu
Sickle-cell hemoglobin
Val
AA
AUG
GT
T
Sickle-cell hemoglobin
Mutant hemoglobin DNAC
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Point Mutations
• Two general categories1. Nucleotide-pair substitutions2. One or more nucleotide-pair insertions or
deletions
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Nucleotide-Pair Substitution
• Replaces one nucleotide pair with another pair of nucleotides
• Silent mutations= no effect on the amino acid because of redundancy in the genetic code
• Missense mutations= codes for incorrect amino acid
• Nonsense mutations= change an amino acid codon into a stop codon– Usually creates nonfunctional protein
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Figure 17.24a
Wild type
DNA template strand
mRNA5
5
Protein
Amino endStopCarboxyl end
33
3
5
Met Lys Phe Gly
A instead of G
(a) Nucleotide-pair substitution: silent
StopMet Lys Phe Gly
U instead of C
A
A
A A
A A A A
A AT
T T T T T
T T TT
C C C C
C
C
G G G G
G
G
A
A A A AG GGU U U U U
53
35A
A A
A A A A
A AT
T T T T T
T T TT
C C C CG G G G
A
A
A G A A A AG GGU U U U U
T
U 35
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Figure 17.24b
Wild type
DNA template strand
mRNA5
5
Protein
Amino endStopCarboxyl end
33
3
5
Met Lys Phe Gly
T instead of C
(a) Nucleotide-pair substitution: missense
StopMet Lys Phe Ser
A instead of G
A
A
A A
A A A A
A AT
T T T T T
T T TT
C C C C
C
C
G G G G
G
G
A
A A A AG GGU U U U U
53
35A
A A
A A A A
A AT
T T T T T
T T TT
C C T CG
G
GA
A G A A A AA GGU U U U U 35
A C
C
G
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Figure 17.24c
Wild type
DNA template strand
mRNA5
5
Protein
Amino endStopCarboxyl end
33
3
5
Met Lys Phe Gly
A instead of T
(a) Nucleotide-pair substitution: nonsense
Met
A
A
A A
A A A A
A AT
T T T T T
T T TT
C C C C
C
C
G G G G
G
G
A
A A A AG GGU U U U U
53
35A
A
A A A A
A AT
T A T T T
T T TTC C C
G
G
GA
A G U A A AGGU U U U U 35
C
C
G
T instead of C
C
GT
U instead of AG
Stop
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Insertions and Deletions• Additions or losses of nucleotide pairs in a gene
– Often has greater negative effect than substitutions• Frameshift mutation• Missing amino acid (loss of 3 nucleotides)
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Figure 17.24d
Wild type
DNA template strand
mRNA5
5
Protein
Amino endStopCarboxyl end
33
3
5
Met Lys Phe Gly
A
A
A A
A A A A
A AT
T T T T T
T T TT
C C C C
C
C
G G G G
G
G
A
A A A AG GGU U U U U
(b) Nucleotide-pair insertion or deletion: frameshift causingimmediate nonsense
Extra A
Extra U53
5
3
3
5
Met
1 nucleotide-pair insertion
Stop
A C A A GT T A TC T A C GT A T AT G T CT GG A T GA
A G U A U AU GAU G U U C
A TA
AG
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Figure 17.24e
DNA template strand
mRNA5
5
Protein
Amino endStopCarboxyl end
33
3
5
Met Lys Phe Gly
A
A
A A
A A A A
A AT
T T T T T
T T TT
C C C C
C
C
G G G G
G
G
A
A A A AG GGU U U U U
(b) Nucleotide-pair insertion or deletion: frameshift causingextensive missense
Wild type
missing
missing
A
U
A A AT T TC C A T TC C G
A AT T TG GA A ATCG G
A G A A GU U U C A AG G U 3
53
35
Met Lys Leu Ala
1 nucleotide-pair deletion
5
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Figure 17.24f
DNA template strand
mRNA5
5
Protein
Amino endStopCarboxyl end
33
3
5
Met Lys Phe Gly
A
A
A A
A A A A
A AT
T T T T T
T T TT
C C C C
C
C
G G G G
G
G
A
A A A AG GGU U U U U
(b) Nucleotide-pair insertion or deletion: no frameshift, but oneamino acid missing
Wild type
AT C A A A A T TC C G
T T C missing
missing
Stop
53
35
35
Met Phe Gly
3 nucleotide-pair deletion
A GU C A AG GU U U U
T GA A AT T TT CG G
A A G
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Accuracy
• Transcription less accurate than replication– Errors occur 1 in 10000 nucleotides
• Protein synthesis more tolerant of errors than replication– Redundancy of codons for amino acids