Final Review Flashcards
What type of cell has a Kozak sequence?
Eukaryotes
What site does the mRNA enter first?
The A site (except for MET and fMET). Just remember the acronym A.P.E. where the tRNA moves from A->P->E, but A is on the right and E is on the left of the ribosome
What protein is bound to tRNA when it is in the cytoplasm?
EFTU hydrolyzes GTP to GDP when the tRNA enters the ribosome, which causes a conformational change to EFTU. This allows EFTU to dissociate and allows the tRNA to fit in the A-site and move through the ribosome
Where does the MET tRNA go first when entering the ribosome?
The P-site.
What is the function of EF-G-GTP in translation?
It powers the ribosome and allows it to ratchet, or translocate, allowing the next mRNA to be read
In eukaryotes what happens first in translation?
The 40S and the MET tRNA move along the mRNA until they find the Kozak sequence
What is mismatch repair?
One base doesn’t match up with the other, the mismatch repair system will cut out the mismatch and replace it with the correct base
How does a Prokaryotic cell recognize a mismatch? How about a Eukaryotic cell?
- -The newly formed prokaryotic DNA strand will be methylated.
- -Newly synthesized eukaryotic DNA strands will have Okazaki fragments (prokaryotes also have these fragments)
What is direct repair and how does it work?
Any type of repair that directly reverses the mutation (Ex. reversal of dimer formation)
What is base excision repair?
The incorrect base/damaged base is cut out, and then a cut is made in the backbone
What is nucleotide excision repair?
When two phosphodiester bonds (on either side of the nucleotide) are cut in the DNA backbone and an entire nucleotide is removed
When does non-homologous chromosome recombination occur?
When the chromosomes are massively damaged the cell will simply put together chromosomes that are nearest each other. This is a last ditch effort to save the genetic information.
Which amino acids are acidic/positively charged and how can you recognize them?
Aspartate and Glutamate
–Look for the carboxylic acid side chain
Which amino acids are basic/negatively charged and how can you recognize them?
Histidine, Lysine, Arginine
–Look for multiple nitrogens or nitrogens at the end of a chain of carbons
Which amino acids are subject to phosphorylation by kinase and why?
- -Serine, Threonine, Tyrosine (Remember PSTT)
- -B/c they contain terminal OH groups
Which amino acid can disrupt an alpha helix?
Proline
–“I am a pro at disrupting the alpha helix”
What amino acid can form di-sulfide bonds?
Cysteine
–Just look for the terminal Sulfur, or think “The Cysteine chapel is a stable structure”
What type of bond stabilizes secondary structures and where does it occur?
Hydrogen bonding between the protein backbone atoms
What does Urea do to proteins?
Breaks non-covalent bonds
What does beta-mercaptoethanol do to proteins?
Breaks disulfide bonds
What is the single molecular difference between a dNTP and an NTP, and where is it located?
There is only a hydrogen attached to the 2’ carbon in dTNP’s (hence deoxyribose)
What is a nucleoside composed of?
Sugar and a base
What is the diameter of the double helix?
2nm
What is the height of one turn of the double helix?
3.4nm
What is the average weight of a nucleotide?
310 daltons
What is an example of a palindrome sequence?
5’-AACGTT-3’
Its compliment 3’-5’ readsTTGCAA, but 5’-3’ it reads AACGTT
What does methylation do to genes?
It silences them
How many types of histones are there?
5 types: H1, H2A, H2B, H3, H4
What is an octomer made of?
2 copies each of: H2A, H2B, H3, and H4
Why is GAPDH used in experiments?
It is the housekeeping gene. It is a loading control to ensure that all samples were not contaminated and that the actual experimental procedure didn’t vary from sample to sample. The GAPDH signal should be comparable across your blot.
Is the sense/coding strand what is read when RNA is made?
No, the template strand is read. This is why the RNA has the same sequence as the coding strand, but with U replacing T.
In which direction do restriction enzymes recognize and cut a recognition sequence?
5’ to 3’
Do bacteria have nucleosomes?
No, but they do have small proteins which aid in condensing their DNA
What is a nucleosome?
An octomer, H1 histone, and associated DNA
–Just remember it is like a spool and thread
What does acetylation do to the histone tails?
It converts them from a positive state to a neutral state (this loosens the DNA. Just remember that DNA is negative and de-acetylated histones are positive)
What is aneuploidy?
Is any abnormal number of chromosomes
What is the polylinker?
A segment of several unique recognition sites for restriction enzymes in a plasmid
What is the difference between stable and transient transfection?
Transfection is the process by which foreign DNA is incorporated into and expressed by native DNA. In transient transfection, the DNA does not integrate with its hosts DNA. In stable transfection it does.
What is significant about cDNA?
It is made from mRNA, and therefore the introns have been removed.