Studies On The Expression And Purified Condition Of Anepâ…¡

There are a lot of polypeptides that have special biological activity in scorpion. ANEPII is one of the most important polypeptides in Buthus martensii Karsh. Transformed the recombinant expression vectors into the E.coli BL21(DE3), and then induced the expression by adding IPTG. In order to improve the expression level of ANEPII in E.coli, growth conditions of the recombatant ,which influence the final yield of protein expression, including inoculation ratio, inducing time and concentration of IPTG and so on, were studied in shaking flasks.The recombinant ANEPII-his6 is electrophoresis purity after Chealting Sepharose Fast Flow , and analyzed by SDS-PAGE.To test the stability of the expression vectorpNJUTRXA-ANEPII, inoculated one time every 12h, after 2,4,6 times, the result was that the mutation rate of the pNJUTRXA-ANEPII is 0.002%, 0.012%, 0.016%;, the losing rate is 1.2%, 4.6%, 7.3%. It indicated that the expression vector have good stability.