The Study To Analysis And Screen Of RhbFGF Prokaryotic High-expression Systems By Computer Assisted

Computer software-assisted analysis is more and more important for the research of Genetics and Molecule Biology to design, screen and assess experiments. To explore high expression prokaryote system of exogenous hbFGF, twenty modified upstream primers and one downstream primer were designed by DNASIS 2.5 software and then were employed to adjust the translation initiation region (TIR) , reduce G+C content and raise A G?through nucleotide change. Using pET-3c as vector, the cloned rhbFGF genes were expressed in BL21(DE3) plysS. Most rhbFGF were expressed in E.coli ≥ 10% of total cellular protein. And 5 recombinants have much soluble rhbFGF. One of these recombinants by random chose was fermented. The target protein from the supernatant of bacteria lysate was purified. The immunology character was detected by antibody in western blot. The bioactivity of the purified rhbFGF was identical with the standard bFGF. The result showed this rhbFGF had the same amino acid sequence, bioactivity and immunology character with nature hbFGF. The conclusion indicted the computer software-assisted analysis in genetics and molecular biology has good prospect.