Tuesday, September 26, 2006

At Least Theyve Got Booze in Casinos...

My job is so damn addicting. Its like playing the slots—you can plug in quarters forever thinking “Maybe this time Ill win! I can feel it! One more spin!” And just when you’re starting to give up—BAM! Jackpot!!! Today was almost a jackpot:



AWWWWWWWWW!! Non-infectious construct! God dammit!

Ugh. 6 months of work. One more week and Ill get that last construct. I can do it.

What youre looking at:
My job is to make various laboratory strains of HIV to help us understand exactly how HIV attaches to our cells, so we can design vaccines in a more logical manner. I take those viruses and infect TZM-bl cells.

TZM-bls are ‘indicator cells.’ That means that some genes have been introduced into their genome-- luciferase and B-galactosidase, attached to the HIV promoter region. If the viruses are infectious, they will infect the TZMs, and bring along with them a gene called ‘tat’, which helps transcribe the infectious viruses genome, and also the artificially inserted genes. You can either do a ‘luciferase assay’, or a ‘beta-gal staining assay’ to see if you got any infected indicator cells. Those blue globs you see in the pics are infected cells.

Another cool thing about these indicator cells is that when they are infected with HIV, they will form ‘syncytia.’ The envelope gene is expressed on the surface of the cells. HIV will normally steal the cells membranes, and their env, when they bud off from the infected cell. When the env on the surface of the cells comes into contact with receptors on uninfected cells, they will fuse together, forming multinucleated blobs!


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