Optimization Expression Of The Recombinant Human HBV-Rnase H In E.coli And Establishment Identification Of Anti HBV-RNaseH McAb Hybridoma Cell-lines

Aims: To optimize the conditions of the recombinant human HBV/RNAse H gene expressed in E .coli BL21 by genetic engineering techniques and to establish hybridoma cell line which can secrete monoclonal antibodies against HBV recombinant RnaseHl and H2 protein. Methods: We analyzed and optimized all kinds of expression conditions, such as the E.coli host cells BL21(DE3) codon plus?-RP, HB101, BL21(DE3)PlysS, GI727DH5α, BL21, culture medium ( LB, SOB, SOC, TB, 2YT, M9 ), induction time (when OD was 0.2, 0.4, 0.6, 0.8, 1.0, Irrespectively) , induction period (2.0h,4.0h,6.0h) , pH (6.0, 6.5, 7.0, 7.5, 8.0, 8.5) and induction concentration of IPTG (0.1, 0.3, 0.6, 1.0, 2.0, 3.0 mmol/L). The stability of the recombinant plasmids...