BIOL 200 MCGILL TEST WITH
COMPLETE SOLUTIONS
RNA polymerase - ANS-Core enzyme + sigma factor = Holoenzyme
Sigma Factor - ANS-Finds promoter (motif rich in T/A)
Core Enzyme - ANS-Two alpha and two beta proteins
Holoenzyme - ANS-Scans DNA to find promoter (help of Sigma factor) forms closed
complex.
Ribosome - ANS-RNA + Protein. Large subunit 50s, small is 30.
Together = 70s
Initiation Factors (3) - ANS-IF1/IF3 recruits 30s ribosome,
IF1/IF2 recruits 50s subunit, forming the 70s
Elongation Factors (Efs) - ANS-to make peptides bonds between aa, ribozyme 23s
rRNA makes reaction
Release factors (RFs) - ANS-RF1/RF2 mimics a tRNA.
RF3 allows for disassembly of 70s ribosome, release of polypeptide.
DNA Helicase - ANS-Seperates the DNA strands
Tropoisomerase - ANS-relieves supercoils when unwinding the DNA
Primase - ANS-RNA polymerase, adds primer
Pol alpha - ANS-DNA polymerase extends the primer with dNTPs for 20 bp (approx)
pol E/Rfc/PCNA complex - ANS-For leading strand
pol epsilon (pol e) - ANS-DNA polymerase which extends the leading strand with
dNTPs.
Rfc (Replication factor C) - ANS-loads/unloads complex on strand.
, what are the components of plasmids? - ANS-ORI: Origin of replication, to be able to
multiply as many times as we need.
ampr : Ampicillin resistance gene.
Polyinker: Region to put the DNA of interest, number of restriction endonuclease sites
that can be recognized by enzymes.
How do polyinkers work? - ANS-Restriction sites (palindromic sequences) where
Endonuclease will cut and create "sticky ends". The genomic DNA of interest will anneal
to the sticky ends of the plasmid. T4 DNA ligase will complete the backbone break.
Results in a Recombinant Plasmid.
What are the steps to transformation? - ANS-1. Mix E.Coli, Plasmid, and CaCl2+ Pulse
of heat.
2. Place the mix on agar plate containing ampicillin. (The bacterias that got transformed
with the plasmid will survive because of ampr, the rest will die)
3. E.Coli with plasmid will colonize, each containing multiple copies of the recombinant
plasmid.
(note that plasmid replication occurs before cell division)
What are the ingredients needed to perform a PCR? - ANS-1. DNA template
2. Taq DNA polymerase (Heat tolerant)
3. Primers (oligos). approx 20 nucleotides long.
4.dNTPs
Steps to PCR - ANS-1. 95 degrees (denature of DNA)
2. 60 degrees, ideal temperature for the primers to anneal. (Note, higher G/C content
will need a higher annealing temperature)
3. 72 degrees, DNA extension with Taq DNA polymerase.
4. Repeat this 20 to 40 times
Reaction mix of DNA sequencing (dideoxy Chain termination Method) - ANS-1. DNA
polymerase
2. Oligonucleotide primers
3. DNA template
4. dNTPs (100 mM)
5. ddNTPs (1 mM)
Genome - ANS-entirety of an organism's heredity info (non-coding and coding DNA)
Coding DNA - ANS-Gene; nucleic sequence necessary for the synthesis of a functional
gene product. (polypeptide, RNA structure)
What is a polycistronic transcript ? - ANS-mRNA encodes for more than one protein.
Mostly in prokaryotes, rare in eukaryotes.
COMPLETE SOLUTIONS
RNA polymerase - ANS-Core enzyme + sigma factor = Holoenzyme
Sigma Factor - ANS-Finds promoter (motif rich in T/A)
Core Enzyme - ANS-Two alpha and two beta proteins
Holoenzyme - ANS-Scans DNA to find promoter (help of Sigma factor) forms closed
complex.
Ribosome - ANS-RNA + Protein. Large subunit 50s, small is 30.
Together = 70s
Initiation Factors (3) - ANS-IF1/IF3 recruits 30s ribosome,
IF1/IF2 recruits 50s subunit, forming the 70s
Elongation Factors (Efs) - ANS-to make peptides bonds between aa, ribozyme 23s
rRNA makes reaction
Release factors (RFs) - ANS-RF1/RF2 mimics a tRNA.
RF3 allows for disassembly of 70s ribosome, release of polypeptide.
DNA Helicase - ANS-Seperates the DNA strands
Tropoisomerase - ANS-relieves supercoils when unwinding the DNA
Primase - ANS-RNA polymerase, adds primer
Pol alpha - ANS-DNA polymerase extends the primer with dNTPs for 20 bp (approx)
pol E/Rfc/PCNA complex - ANS-For leading strand
pol epsilon (pol e) - ANS-DNA polymerase which extends the leading strand with
dNTPs.
Rfc (Replication factor C) - ANS-loads/unloads complex on strand.
, what are the components of plasmids? - ANS-ORI: Origin of replication, to be able to
multiply as many times as we need.
ampr : Ampicillin resistance gene.
Polyinker: Region to put the DNA of interest, number of restriction endonuclease sites
that can be recognized by enzymes.
How do polyinkers work? - ANS-Restriction sites (palindromic sequences) where
Endonuclease will cut and create "sticky ends". The genomic DNA of interest will anneal
to the sticky ends of the plasmid. T4 DNA ligase will complete the backbone break.
Results in a Recombinant Plasmid.
What are the steps to transformation? - ANS-1. Mix E.Coli, Plasmid, and CaCl2+ Pulse
of heat.
2. Place the mix on agar plate containing ampicillin. (The bacterias that got transformed
with the plasmid will survive because of ampr, the rest will die)
3. E.Coli with plasmid will colonize, each containing multiple copies of the recombinant
plasmid.
(note that plasmid replication occurs before cell division)
What are the ingredients needed to perform a PCR? - ANS-1. DNA template
2. Taq DNA polymerase (Heat tolerant)
3. Primers (oligos). approx 20 nucleotides long.
4.dNTPs
Steps to PCR - ANS-1. 95 degrees (denature of DNA)
2. 60 degrees, ideal temperature for the primers to anneal. (Note, higher G/C content
will need a higher annealing temperature)
3. 72 degrees, DNA extension with Taq DNA polymerase.
4. Repeat this 20 to 40 times
Reaction mix of DNA sequencing (dideoxy Chain termination Method) - ANS-1. DNA
polymerase
2. Oligonucleotide primers
3. DNA template
4. dNTPs (100 mM)
5. ddNTPs (1 mM)
Genome - ANS-entirety of an organism's heredity info (non-coding and coding DNA)
Coding DNA - ANS-Gene; nucleic sequence necessary for the synthesis of a functional
gene product. (polypeptide, RNA structure)
What is a polycistronic transcript ? - ANS-mRNA encodes for more than one protein.
Mostly in prokaryotes, rare in eukaryotes.
