9899 Final

Section I:

(Prof. Lam’s Portion; 30 marks total)

Question 1: Answer ALL parts. Each question carries 3.5 marks. Briefly describe the principles and major applications of the following techniques: (a) (b) (c) (d)

Oocyte microinjection. Electroporation Retrovirus-mediated transformation Yeast functional complementation

Question 2: Read the whole question and attempt ALL parts. Even if you do not know the answer of some parts, you may still be able to answer the rest. Experimental details are not required. Just describe the underlying principles. The expression of Gene A in the model plant Arabidopsis thaliana is higher in light and low in dark conditions. Both the promoter and structural region of Gene A were cloned and sequenced. The mRNA and protein product of Gene A are, however, very difficult to detected. (a)

Design a transient plant expression system to test whether the light induction is at the transcriptional or the translational level. (4 marks)

The regulation of Gene A was subsequently found to be at the transcriptional level. The promoter of Gene A is located in a 2 Kb region upstream from the transcriptional start site. Further experiments also demonstrated that the promoter region between +1 to –100 (base pairs 5’ of the transcriptional start site) is essential for transcription but is not related to the light induction. Based on the DNA sequence information, unique restriction sites flanking the regions –100 and –2000 (base pairs 5’ of the transcriptional start site) were identified. (b)

Using the same system you proposed in (a), design an experiment to locate the approximate position of the cis-element in the promoter that is responsible for the light induction. (4 marks)

It is suspected that the light induction of Gene A is mediated by the transcription factor Protein X (encoded by Gene X). The cDNA of Gene X was cloned and sequenced. (c)

Using the same system you proposed in (a), design an experiment to test whether Protein X is involved in the light induction of Gene X. (4 marks)

It is also suspected that Protein X will form a dimer (i.e. two molecules of Protein X to link noncovalently to form one functional transcription factor). However, the region responsible for the dimerization is unknown. (d)

Design an experiment to demonstrate that Protein X will form a dimer. (4 marks)

Section II:

(Prof. Sun’s Portion; 22 marks total)

Answer ALL questions 1. You are the laboratory director of a national plant molecular biology institute involving in genome research. Recently the institute was given the responsibility to coordinate the soybean genome project among the Hong Kong and China institutes. The goals of the project are to map and sequence the whole soybean genome (20 pairs of chromosomes; 1x108 bp of genome size) in 12 years with a specified (adequate) amount of funding. Your were appointed as the project coordinator and needed to develop a strategic research plan to effectively and efficiently coordinate and carry out this grand project. Outline your plan. (7 marks) 2.

In 1998, a new NIH-DOE 5-year human genome project plan “New goals for the US human genome project: 1998-2003” was developed and published in Science 282:682689 (1998) by Francis S. Collins and other members of the planning group. Among the 8 goals, Goal 1 – The Human DNA Sequence listed and described several sub-goals. Which of these sub-goals made a strong impression on you and why? (3 marks)

3. Biotechnology offers opportunities for innovation in health care. Give three modern medicine examples that you find them innovative. (5 marks) 4. Conventional breeding method involves crossing and back crossing of plants to transfer desirable genes; it thus takes time to develop an improved plant variety. Biotechnological approach is capable of transferring specific genes into a plant through transformation; so it is more precise, direct and quicker in generating new varieties. However, it has been over 20 years since the inception of biotechnology yet only a few genetically engineered products are on the markets. Why takes so long for the development ? What possible reasons you can think of? (5 marks) 5. What are biological and physical containments? Why we need both of them? (2 marks) - END -