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(next week)
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221.S16
Week of MAR-14
M T W R F
10-11 10-11
Iryna Run
office hours office hours
LLAB 114 LLAB 114
11:00-11:50 noon-1:00 11:00-11:50 11:00-11:50
Class GG Class Class
LLAB 10 office hours LLAB 10 LLAB 10
Questions? LL114
LLAB 319
12-1
Steve
office hours
LLAB 114
5-6:30 6:30-8:00 6:30-8:30 5:15-6:15
Review Review Exam-2 GG
LLAB 10 LLAB 10 LLAB 10 office hours
LLAB 319
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221.S16
What is on Exam-2?
Content
• The primary emphasis for the exam will be the material that we covered in
class from Monday 2/8/16 (Allele interactions) through Wednesday 3/2/16
(RNA splicing).
• You are also responsible for the corresponding reading in the text (see
syllabus) and for the required reading listed on our Canvas site.
• You should also have a good understanding of the material & problems
corresponding to the prerequisites for the course (see syllabus).
• As you might imagine, you should be pretty facile at solving problems like
those included on the syllabus and in the recitations.
Review sessions
• Monday 3/14 5:00-6:30 LLAB 10
• Tuesday 3/15 6:30-8:00 LLAB 10
Exam-1
• Wednesday 3/16 6:30-8:30 LLAB 10
• Simple calculators are permitted for this exam. Laptops, iPads, PDAs,
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smartphones, etc. are NOT permitted
221.S16
Clarification
Shown below is a 50-nucleotide double-stranded DNA segment from a prokaryotic genome that contains a
factor-independent transcriptional terminator element.
5’-...GCTTTACAAAAAAAGTGCGCTGCTGCTTAGCAGCGCACACCTTGGTGGGG...
3’-...CGAAATGTTTTTTTCACGCGACGACGAATCGTCGCGTGTGGAACCACCCC...
a. Write the nucleotide sequence of the transcribed RNA. There is no need to indicate any secondary
structure but clearly label the 5’- and 3’-ends of this RNA.
b. If the blue-shaded 3-bp sequence were inverted, would this affect the efficiency of transcriptional
termination? Briefly explain.
c. If the red-shaded 2-bp sequence were deleted, would this affect the efficiency of transcriptional
termination? Briefly explain.
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221.S16
Anatomy of an mRNA transcript
H2N
COOH
5
[UTR: untranslated region]
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Translational initiation: Prokaryotes
3’
5’
formylmethionine
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221.S16 B. Keith
Prokaryotic translation: linear polysomes
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221.S16
Translational initiation in eukaryotes: mRNA binding and scanning
(RBS)
scan->
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221.S16
Translational initiation in eukaryotes: mRNA binding and scanning
The polypeptide chain is
begun with a methionine
attached to a special
initiator tRNAi, which is the
only tRNA that can bind the
ribosomes in the absence of
the 60S subunit…
A complex containing
initiation factors (eIF2, etc.),
the 40S subunit, and Met-
tRNAi binds the m7G 5’ cap, scan->
then scans along the mRNA
looking for the first AUG
sequence it can find…
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221.S16 B. Keith
Translational initiation in eukaryotes: mRNA binding and scanning
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221.S16 B. Keith
Translational initiation in eukaryotes:
Two features recognized by ribosomes
Kozak sequence
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221.S16
Translational termination: protein factors
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221.S16 B. Keith
Macromolecular mimicry: tRNA and Relaese Factors
tRNA
yeast RRF
anticodon
loop
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221.S16 Selmer et al. Science (1999) 286:2349-2352
Properties of the Genetic Code
Non-overlapping code
No punctuation
3-nucleotide codons (triplet code)
Degenerate code
Start & stop codons
Nearly universal
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221.S16
Genetic code
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Mutations within coding regions
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221.S16
Anatomy of an mRNA transcript
H2N
COOH
18
[UTR: untranslated region]
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1 sequence and 3 reading frames
RNA 5’- CAU CAU CAU CAU CAU CAU CAU CAU frame 1
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Translation: reading frames
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221.S16 Figure 8-52a Molecular Biology of the Cell (© Garland Science 2008)
Translation: Open Reading Frames (ORFs)