Experimental Study On Improving The Effect Of BCG Immunized Mice With DNA-85B

Objectives: 1. to get plenty of superpure reconstruct plasmid of PTB-30m. 2. to determine whether the prime–boost immune strategy with DNA-85B and BCG can induce special immune response in mice surpass BCG alone .Methods 1.transformed a little reconstruct plasmid of PTB-30m into E.coli strain of DH5α; selected the strain with ampicillin on LB plates;amplified it in LB liquid culture with ampicillin;extracted the plasmid with endofree qiagin-mega-kit;so we got the ultrapure plasmid of PTB-30m (detect the A260 or A260/A280 with Uvspectrophotometry); then digested it with HindⅢand EcoreⅠ,analyzed it on a 1% agarose gel electrophoresis (170volts,20min).2. C57BL/6 mice were immunized with reconstruct plasmid PTB-30m firstly, then immunized with BCG two weeks later.killed the immunized mice four and eight weeks after the second immunization respectively, prepared spleen lymphocytes, cultured with PPD in vitro. Lymphocyte transformation test was detected by MTT...