Explain how the Harrison group ensured that the RT to DNA tethering reaction would work.

5. Why did Eddie Arnold’s group fail to resolve the issue of understanding the mode of resistance caused by mutations within the b3-b4 loop?

6. Explain how the Harrison group ensured that the RT to DNA tethering reaction would work. Explain your answer. (

7. Explain, in detail, how the researcher was able to determine that the b3-b4 loop rotated toward the palm subdomain of HIV-1 RT

8. Explain why Huang’s paper indicated how ddI, a dideoxy analog to RT, resistance could occur from the mutation L74V.

1. You have cloned RT into an expression vector and you have transformed it into CD4+ T cells. However, you only made a recombinant clone of the p66 subunit. After isolating the protein you have discovered that your protein is made up of the p66/p51 heterodimer. How is this possible if you only cloned the p66 subunit?

2. You want to clone the cytokine IL-12 (insert size 350bp) from mice so that you can compare functions with that of the human IL-12 protein. You have used the vector below and cut the plasmid with the restriction enzyme Kpn I. After transformation and selection on selective media, you isolate the plasmid DNA on an agarose gel. you obtain the results below. After running the gel you decide to continue on with protein isolation. From running a protein gel you have the correct size, but you get no functional protein back though over 50% of the total protein is at the same molecular weight as your protein. (i) What happened to your clone that would explain these results; (Hint, it’s not a mutation); (ii) how would you fix the problem?

3. AIDS dementia is an end-stage symptom pathology in AIDS patients with death being eminent in the near future. Before drug therapy was established, Industrialized Nations saw high AIDS dementia rates in patients who soon succumbed to opportunistic infections. However, India, with an AIDS rate that was 3x higher than the U.S and Europe had very small percentages of AIDS dementia cases. What is your reasoning behind this scenario?

4. You have engineered an expression vector that contains a protein that will inhibit the proper formation of an eyeball in cave fish. However, for this to work, you need to express this protein only in the eye and nowhere else in the fish. Explain how you would go about ensuring that this inhibiting protein, cloned into your vector, will only be expressed within the developing eyeball of the cavefish. You do not need to tell me how you would get the plasmid into the fish itself. just work with the plasmid. (Note: Any sequences you need can easily be obtained from a DNA sequencing database and you can amplify and any sequences via PCR). No need for controls, just explain what you would have to do.