Cloning and high-level expression of scorpion toxin BmKITa1 in Escherichia coli and insect cells.
BmK ITa1 cDNA was cloned and highly expressed in E. coli and insect cell. SDS-PAGE and western blot analysis revealed that subunit molecular weight of expression products is about 40 kDa and 10 kDa respectively. The expression product purified by a Ni(2+)-IDA-sepharose 6B column was toxic for insect, which indicated that it was biologically activity. Furthermore, Quantitative estimation show that the biological activity of recombinant BmK ITa1 from Tn cells was more powerful than from E. coli.
Liu
Zhen
Z
Shanghai Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, P.R. China.
Yang
Guan-Zhen
GZ
Chi
Cheng-Wu
CW
Wu
Xiang-Fu
XF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D008969:Molecular Sequence Data; D009036:Moths; D012604:Scorpion Venoms / Q000235:genetics*
C471081:BmK Ita1 toxin, Buthus martensii; D018076:DNA, Complementary; D012604:Scorpion Venoms