Expression of the APLA(2) Gene from Agkistrodon halys Pallas.
The APLA(2) gene from Agkistrodon halys Pallas has been cloned into the expression plasmid pBLMVL2 and expressed in E. coli RR1. The molecular weight of the expressed product is approximately 14 kD as shown by SDS-PAGE, its expression level is about 30% of the total cellular proteins. The protein was produced as insoluble inclusion bodies. After partially purified by washing the inclusion bodies, the product was denatured and refolded into active form. Then, the expressed APLA(2) was purified by FPLC Superose (TM) 12 and was a single band as shown by SDS-PAGE. The purified expressed protein had specific activity as the native enzyme and cross-reacted with antisera prepared against the native enzyme. The successful expression of the APLA(2) gene from Agkistrodon halys Pallas provides a good basis for further structure-function studies.
Liu
Xiao-Long
XL
Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China. zhouyc@sunm.shcnc.ac.cn
Pan
Hua
H
Yang
Guang-Zhen
GZ
Wu
Xiang-Fu
XF
Zhou
Yuan-Cong
YC
Cai
Ming-De
MD