Study Of The Fermentation Conditions Of Trichosanthin Gene Engineering Strain And Detection Of The Activity Of The Expression On Suppressing Human Immunodeficiency Virus 1 And Herpes Simplex Virus Type 1 And 2

Hybridoma cell lines capable of secreting monoclonal antibodies (MAbs) against Trichosanthin were obtained by fusing mouse myeloma cells (SP2/0) with spleen cells taken from the BAL B/C having been immunized by the Trichosanthin. The MAbs obtained from the Hybridoma cell could specifically react with Trichosanthin. This research may set good foundation for further research on TCS. Then engineering Strain(T239) was cultivated under different conditions in order to find the optimal fermentations that would lead to maximal fusion protein yield. The strain was cultured under different temperature, PH, rotating rate, time, and capability of bottle and results showed that the proper fermentation conditions consisted of: 30°C , PH7.3, 200r/min, 50ml/500mlculture bottle, 30h on rotating cultivation bed, with 2.0g/L glucoseon added. The activity on anti-Human Immunodeficiency Virus 1(HIV-1 )of the expression (Trichosanthin) were detected. The expression were put into the cells infected by HIV-1 and the activity of HIV p24 antigen and RT activity were detected. Results showed that the cells added into trichosanthin showed RT activity negative, p24 Ag negative and no representative cell pathomorphology change while the cells added into negative control showed RT activity positive, p24 Ag positive and representative cell pathomorphology change, and the cells added into zidovudineshowed RT activity negative, p24 Ag negative and no representative cell pathomorphology change. The expression can inhibit Human Immunodeficiency Virus 1 in vitro. To detect the anti-Herpes Simplex Virus typel and 2 activity of trichosanthin expressed by the means of biotechnology, the trichosanthin were put into the cells infected by HSV-1 and HSV-2 to see if it can suppress HSV-1 and HSV-2 or not. The cells having trichosanthin showed ELISA negative and no representative cell pathomorphology change while the cells having not trichosanthin showed ELISA positive and representative cell pathomorphology change. Trichosanthin can also suppress HSV-1 and HSV-2 effectively in vitro.