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Name: Student ID

Molecular Biology and Genetic Engineering (BIO4320) Feb. 26, 1999


Mid-term Examination (Total 48 marks)

Part I (Dr. Fung’s Part; 16 marks)

1. Name three factors that will affect the forms of the DNA structure. (1.5 marks)

2. Write down all the codons that can be paired by the tRNA with an anti-codon 5’-ICC-3’. (1.5
marks)

3. Draw a diagram to illustrate the 3 different types of eukaryotic promoters that are recognized
by the RNA polymerase I, II and III. Name one gene product that can be transcribed by each
of these 3 different RNA polymerase. (3 marks)
Name: Student ID

4. Draw a dia gram to illustrate the structure of a typical eukaryotic gene. (2 marks)

5. The bacterial cells were first cultured in a glucose medium and then shifted to a medium
contains lactose as the only carbon source. Describe the changes of the gene expression that
the E. coli cells required to cope with the changes in the nutrient supply. (8 marks)

- END of Part I -
Name: Student ID

Molecular Biology and Genetic Engineering (BIO4320) Feb. 26, 1999


Mid-term Examination (Total 48 marks)

Part II (Dr. Lam’s Part; 32 marks)

(1) Distinguish the following terms: (3 marks each)

(a) Lambda insertion vector and replacement vector

(b) Phagemid and cosmid

(c) Isoschizomers and compatible ends

(d) Competitive and non-competitive internal standards for quantitative PCR.


Name: Student ID

(2) In Year 2100, human in the Earth (Human-E) has established scientific exchange programs
with the highly intelligent human-like living organism in Mars (Human-M). Dr. Lam’s
undergraduate students taking the BIO4320 course were asked to study the genetic homology
between Human-E and Human-M.

(a) In the first experiment, the students were asked to clone and amplify a specific gene (Gene X)
from Human-M and express the encoding protein (Protein X) in a yeast system for further
analysis. Design a plasmid vector which can serve this purpose. You should describe all the
necessary features of this plasmid vector. (8 marks)
Name: Student ID

(b) Protein X from Human-M was later found to be consisted of two functional domains (I and II)
and a linker region (L). A similar protein (Protein X’) found in Human-E also consisted of
two functional domains (I’ and II’) and a linker region (L’) (see diagram below).

Domain I L Domain II Protein X

Domain I’ L’ Domain II’ Protein X’

Dr. Lam asked his students to swap the functional domains and create a chimeric protein with
Domain I and Domain II’ as shown below:

Domain I Domain II’ Chimeric Protein

The students decided to using recombinant DNA techniques to solve this problem. They
sequenced the linker region and found some unique restriction sites that did not occur in any
of the functional domains. The DNA sequence of the linker region and the amino acids
encoded were shown below:

5’...........TTT CTG CAG GGG AAA...........3’


3’...........AAA GAC GTC CCC TTT...........5’ Protein X

[Domain I].. Phe Leu Gln Gly Lys..[Domain II]

5’............TTT CCC GGG ATC CAA............3’


Protein X’
3’............AAA GGG CCC TAG GTT............5’

[Domain I’].. Phe Pro Gly Ile Gln..[Domain II’]

The students were provided the following enzymes but without any linkers and adapters.

Klenow; T4 DNA polymerase; E. coli ligase; T4 DNA ligase

BamHI 5’ GGATCC 3’
3’ CCTAGG 5’

PstI 5’ CTGCAG 3’
3’ GACGTC 5’

SmaI 5’ CCCGGG 3’
3’ GGGCCC 5’

XmaI 5’ CCCGGG 3’
3’ GGGCCC 5’
Name: Student ID

(c) [continued]

Based on the information and materials provided above, describe how the chimeric DNA
construct encoding a chimeric protein (with domain I and II’) can be made. You should
explain the enzymes involved in each step of the construction. (12 marks)

Remark: think about it carefully, only certain enzymes work.

-END of Part II-