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BCH5413 Exam One 2024 with Questions and 100% Correct Answers

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What is the difference between and intron and an exon? - Answer Introns are non- coding sections and exons are coding sequences. How can one stretch of DNA be used to create more than one protein? - Answer It can undergo alternative splicing and create different protein products. What is the me...

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BCH5413 Exam One 2024 with Questions and
100% Correct Answers

What is the difference between and intron and an exon? - Answer Introns are non-
coding sections and exons are coding sequences.

How can one stretch of DNA be used to create more than one protein? - Answer It can
undergo alternative splicing and create different protein products.

What is the meaning of the term exon shuffling? - Answer Exon shuffling is when an
exons from different genes are brought together or the same exon is duplicated.

How is this process related to the development of new proteins? - Answer This process
is essential for the development of new proteins because it alters the existing structure
and allows the exons to code for a new protein.

After isolating intact chromatin from cells, lightly digesting it with micrococcal nuclease
and running it on an agarose gel, you see a banding pattern of repeating 200bp
fragments (200, 400, 600 etc). Explain this result. - Answer This is because part of the
DNA is protected from degradation by the nuclease by the nucleosome.

Explain the role of each of the histones in the formation of chromatin? - Answer
Histones interact with each of the 14 minor grooves of DNA and the DNA wraps around
them.
-H4/H2a/2b/H3 (core) are all wound up with DNA and they help expose the DNA for
transcription.
-H1 leads to more complex nucleosomal DNA. Ultimately will result in up-down
structure where it is very organizes and DNA is 7 fold more compact where it can be
inside the nucleus. H1 leads to next level of chromatin structure.

,T/F Blunt ends gives better ligation than sticky ends - Answer False, sticky ends do
better because they give a good place to bind in and ligate

T/F The general ratio of vector to insert in cloning is 1:3 - Answer True

T/F If bacterial cells have taken up the plasmid, the colonies are white in blue/white
selection. - Answer False, If it has taken up the plasmid the colonies are blue

T/F E.coli methylate their own DNA to prevent to prevent infection by viruses - Answer
True

Why would it be advantageous for a plasmid vector to have each of the following
features:
polylinker or multiple cloning site - Answer They make it easier for DNA cleavage to
occur because of multiple sites for restriction enzymes.

Why would it be advantageous for a plasmid vector to have each of the following
features: antibiotic resistance gene - Answer It confers a protective advantage. Selective
marker

Why would it be advantageous for a plasmid vector to have each of the following
features: origin of replication - Answer Specific sequence that bacterial host recognizes
and will bind to it and copy the plasmid into the host cell

Why would it be advantageous for a plasmid vector to have each of the following
features: LacI - Answer It is used as regulation for LacZ

, Why would it be advantageous for a plasmid vector to have each of the following
features: LacZ - Answer Makes it easy to detect the plasmid through blue/white test

In a cloning experiment, how do you get rid of the bacteria that do not take up the
plasmid? - Answer You can grow them on a plate with ampicillin and the ones that do
not get the plasmid will die on the plate because plasmid confers ampicillin resistance.

What is the function of IPTG in cloning? - Answer Induces lac operon and the cell will
make protein for you

in a cloning experiment, what does it mean when you get colonies on a No ligase / No
insert control plate? - Answer This means that your plasmid was either incompletely cut
by the endonuclease or it somehow recircularized itself.

What if there are more colonies there than on your experimental? - Answer Thats not
good if there are more there than on your experimental because likely you don't have
any samples with you plasmid in it.

In what two ways did the development of the TOPO cloning vector make cloning easier?
- Answer One step combination of the plasmid and insert (taq pcr product) and
topoisomerase will link the DNA insert into a region with the multiple cloning site.

What is a shuttle vector? What additional features (beyond #2) would you want in a
shuttle vector? Why? - Answer Do your cloning in e.coli then shuttle it into a different
host cell system. Also want yeast origin and centromere so that the other host system
can recognize the plasmid and replicate it. Also want a selective marker for the yeast.

In creating a cDNA library what is the advantage of using beads containing oligo T
nuceotides for isolating the RNA? - Answer Used to isolate mRNAs because they will
anneal to the poly a tail

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