Enzyme Specific Antibody Production Using Salmonella typhimurium Proteins that are Explicitly Expressed in Cytoplasmic or Periplasmic Regions
Garrett, Jasmine 1,2 and Curtiss III, Roy2
1School of Life Sciences, Arizona State University; 2Center for Infectious Diseases and Vaccinology, The Biodesign Institute at Arizona State University
Our focus is to develop antibodies that specifically recognize cytoplasmic or periplasmic proteins from Salmonella typhimurium. The cytoplasmic mutL and periplasmic Mbp proteins of Salmonella typhimurium will be selected for this purpose. Mbp and MutL genes will be amplified from Salmonella typhimurim and cloned into a pBAD/His vector. The recombinant plasmid will be introduced into E. coli Top10 by transformation and positive colonies will be selected using ampicillin. After confirming the sequence of the cloned genes, the His-tagged MutL and Mbp proteins will be purified on a Nickel column. The purified proteins will be administered to 6 week old rabbits for antibody production. The serum from the immunized rabbits will be tested for anti-MutL and anti-Mbp antibodies by western blotting using purified proteins and Salmonella typhimurium cell lysate.