BIOT643-UMUC- Summer 2016

Midterm Exam (150 points)

Due in your assignment folder by midnight Sunday July 3.

Based on honor code - Answers provided must be in your own words.

Part I: Multiple choice Questions: Choose the most appropriate answer.

(5 points each).
1. The abundance of mRNA in mammalian cells is measured to be:
a) 1% of total RNA content
b) 10% of total cell content
c) 80% of total RNA content
d) The same as protein content
2. RNAi is used for one of the following purposes.

a) To disrupt a DNA
b) To clone a particular gene
c) To verify the function of a gene
d) To produce a new gene

3. Advantages of fusion proteins can include:

a) Facilitate purification
b) Enhance solubility
c) Increase stability
d) All of the above

4. Bacteria are unsuitable for expression of some eukaryotic proteins, because:

a) They lack a nuclear membrane

b) They have a single chromosome
c) They dont perform all post-translational modifications
d) They produce unsuitably stable proteins

5. To successfully express a protein from a eukaryotic expression vector, the

following is
NOT required.

a) Selectable marker
b) Ribosome binding site
c) Strong eukaryotic promoter
d) Termination sign
6. Which of the following methods is NOT commonly used for high throughput gene
expression analysis?

a) Differential Display analysis

b) SAGE
c) DNA microarray
d) Northern blot hybridization

7. Why would a lab technician choose a bacterial artificial chromosome for cloning a gene,
rather than a plasmid?

a) Because the DNA insert is from genomic DNA and not plasmid DNA
b) Because the insert to be cloned is very large.
c) Because bacterial DNA will be used to make many copies of the gene (i.e., clone it).
d) Because the gene insert will eventually be used to transform cells, meaning
incorporate the gene into genomic DNA.

8. How can both Southern and Western blotting assist with genetic recombination
experiments?

a) Both methods will confirm that the gene is in the plasmid before it is transfected into
cells.
b) One will identify the specific DNA in the plasmid, and the other will identify the specific
protein part of the plasmid.
c) One is used to recombine the plasmid and the other is used to identify the gene
sequences in the plasmid.
d) One will identify a specific DNA sequence in transformed cells and one will identify
expression of the protein from that sequence.

9. Which statement is false regarding DNA libraries?

a) A library is made using several different endonucleases.

b) Each DNA fragment is cloned into a different bacterial cell.
c) DNA sequences from the library are separated by base pair length before cloning into
bacteria.
d) The same gene can be present in fragments of different base pair length.
e) A library is the sum of all sequences in genomic DNA.

10. Which of the following steps is inhibited during the process of RNA interference?

a) DNA replication
b) Transcription
c) Translation
d )Post translational modification
11. Which one of the following techniques is commonly used to quantify the level of mRNA in
a sample tissue?

a) Real time RT-PCR

b) Western blot
c) Far Western Blot
d) Southern Blot

12. Which one of the following methods cannot be used to analyze differential gene (mRNA)
expression?

a) Western Blot analysis

b) Microarrays

c) SAGE

d) Suppression Subtractive Hybridization

Part II: Short answer questions

Please answer the following questions in about 100-200 words. (10 points each)

1. If you want to identify genes from human liver that are highly expressed only under
conditions of high stress, what kind of DNA library will be the best one to use and why?

2. A. What are fusion proteins? Provide a brief description of steps involved in producing and
purification of fusion proteins and two main advantages of using fusion proteins.

3. For gene expression analyses, cDNA is used both in DNA microarray and SAGE methods.
Explain how and why cDNA is used in each method.

4. What are promoters? Why a strong and regulated bacterial promoter is required for protein
expression?

Part III: Long answer question

Please answer the following questions using about 500 words each. For these
questions you must demonstrate your knowledge and understanding of the
techniques presented in lectures and in class readings. Be sure to provide a detailed
answer and proper citation. (25 points)

19. In recent studies, a direct correlation was found between too much sitting and the
development of various health conditions, such as diabetes, cardiovascular disease, and
some types of cancer. As the leading scientist in a related project, you must design a series
of experiments that allows you to identify the gene(s) that play a role in the development of
cancer. In your design provide an outline of the appropriate techniques and methods used
for:
1) DNA library construction
2) Gene expression analysis
3) Target validation/functional characterization
4) Protein expression, including your choice of protein expression system

and any additional information that might be helpful. You should also include the reasons
for choosing your specific approach. Feel free to use schematics to help with your answer.

20. Read the article at http://www.pnas.org/content/94/23/12457.full and answer the following

questions.

1) What approach/methodology did scientists use to determine the role of myostatin in

muscle regulation? Based on their results explain the mechanism of action of the myostatin
gene? (10 points)

2) Knowing what the authors of this study did and what they achieved, explain in sufficient
detail one approach discussed in class, by which you can address the following problem:

Suppose, all you have in your lab is the DNA sequence of the myostatin gene. Using
only that information, devise a way (not the one described in the article) to create strong
muscles in a person suffering from a weak muscle disorder. (15 points)