High Expression of a cDNA Coding for Human Lymphotoxin alpha Derivative in E. coli and Purification of Its Protein Product.
Using PCR, a cDNA coding for human lymphotoxin alpha derivative (hLTalphaDa) lacking 27 amino acids at N-terminal of natural hLT was constructed. The expression construct was expressed in E. coli BL21 (DE3). The product of expression was in the form of inclusion bodies, and accounted for 60%-80% of total bacterial proteins. The product protein was purified to over 95% by treatments of inclusion bodies. The specific activity of hLTalphaDa was above 2x10(8) IU(per mg protein). Its cytotoxic activity can be neutralized by monoclone antibody against hLT. The anti-tumor effects of hLTalphaDa were also tested in vitro and in vivo.
Li
Ping
P
Institute of Genetics, State Key Laboratory of Genetic Engineering, Fudan University, Shanghai 200433, China. changbenli@fudan.edu.cn
Yu
Hong
H
Ni
Zhuo-Yu
ZY
Chen
Chun-Mei
CM
Zhao
Shou-Yuan
SY
Li
Chang-Ben
CB