First Cloning Experiment Cohen et al., 1973 “Recombinant DNA”

Cloning Vectors: pBR322 plasmid Cloning into pBR322

RT-PCR and Cloning

Palmiter et al. (1982) Dramatic growth of mice that develop from eggs microinjected with metallothionine-growth hormone fusion genes. Nature 300: 611-615 “Supermice”

Gene Therapy: Adenovirus vector

Golden Rice

First Glowing Pet Weeding Out Land Mines

Antibody Production Y Polypeptide “X” Anti-”X” Antibodies Antibodies can be utilized to detect the presence of protein “X” in a given sample Y Antibody production Next, I wanted to determine if ribosomes from different tissues are different with respect to P-protein composition. The first thing I needed was a way to detect the individual P-proteins in a ribosome sample and the way I did this was through the use of antibodies. Antibodies are proteins produced by an animals immune system in response to a foreign substance. The antibodies bind to the foreign substance and “tag” them for distruction. The antibodies are specific for that foreign substance and this feacture is what makes them useful to scientist. We can inject a polypeptide into rabbit, the rabiit’s immune system recognizes it as foreign and makes antibodies against that protein. We can then collect the blood from the rabbit, which will contain our anti-X antibodies. Now these antibodies can be utilized to detect the presence of protein X in a given sample Anti-”X” Antibodies

Incubate with Antisera Transfer to nitrocellulose Western Blot Analysis Isolate Proteins SDS-PAGE S6 Y Incubate with Antisera Transfer to nitrocellulose Y P3 And this is a procedure where proteins, ribosomes in my case, are fractionated through a denaturing polyacrylamide gel, or matrix. This is a process which then separates the proteins based on size. Once the proteins are run through the gel, they are transferred to a nitrocellulose membrane to immobilize them. The membrane is then incubated with the P-protein specific antisera and then we go through a final step where the antibodies are detected.

Proteomics Approach in Arabidopsis