Research Details
Out of couch at 3:15, early morning spin to Portland International, crash at my parents' house for a few hours of much need zombie protection, off to OSU for some lab work - the story of my morning.
Research has picked up a bit. Yesterday I received five agar plugs of bacteria in the mail. Two were Escherichia coli containing plasmids (circular, non-genomic pieces of DNA) which I'll call A and B. The plan is to chop out part of plasmid A using restriction enzymes (DNA scissors) and add it to plasmid B. We plan to clone viral genes into the modified plasmid B, and the DNA sequences in the added piece of DNA from plasmid A will enable our virus genes to be transcribed and the eventual protein to go to the outside of the bacterium. It is important to have the protein on the bacterial surface so the horse can mount an immune response against it.
We plan to initially use E.coli (we got some plain old E.coli as our third agar plug) to stick our modified plasmid into, and then move on to Listeria monocytogenes once we have the virus gene(s) in the plasmid(s). L. monocytogenes (bacterium in the fourth and fifth agar plugs) is the bacterium that will finally be used as the vector and immune-stimulant for the projected virus DNA vaccine. The two L. monocytogenes strains we received have genes knocked out of them in order to decrease their pathogenicities. L. monocytogenes can cause a pretty awful illness with an overall death rate of 20% in humans, so the decreased pathogenicity is a very good thing.
Today I received the pieces of DNA (primers, or oligonucleotides) I ordered on Wednesday. I set up two polymerase chain reactions with them to amplify the E5 and E6 genes from bovine papilloma virus 1. Here is an animation of the polymerase chain reaction and here is a text description of the process.
"I'm a goin' home..." (ref. New World Symphony by Antonin Dvorak)
Research has picked up a bit. Yesterday I received five agar plugs of bacteria in the mail. Two were Escherichia coli containing plasmids (circular, non-genomic pieces of DNA) which I'll call A and B. The plan is to chop out part of plasmid A using restriction enzymes (DNA scissors) and add it to plasmid B. We plan to clone viral genes into the modified plasmid B, and the DNA sequences in the added piece of DNA from plasmid A will enable our virus genes to be transcribed and the eventual protein to go to the outside of the bacterium. It is important to have the protein on the bacterial surface so the horse can mount an immune response against it.
We plan to initially use E.coli (we got some plain old E.coli as our third agar plug) to stick our modified plasmid into, and then move on to Listeria monocytogenes once we have the virus gene(s) in the plasmid(s). L. monocytogenes (bacterium in the fourth and fifth agar plugs) is the bacterium that will finally be used as the vector and immune-stimulant for the projected virus DNA vaccine. The two L. monocytogenes strains we received have genes knocked out of them in order to decrease their pathogenicities. L. monocytogenes can cause a pretty awful illness with an overall death rate of 20% in humans, so the decreased pathogenicity is a very good thing.
Today I received the pieces of DNA (primers, or oligonucleotides) I ordered on Wednesday. I set up two polymerase chain reactions with them to amplify the E5 and E6 genes from bovine papilloma virus 1. Here is an animation of the polymerase chain reaction and here is a text description of the process.
"I'm a goin' home..." (ref. New World Symphony by Antonin Dvorak)
posted by Claire at 17:09
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