[Expression and bioactivity analysis of staphylococcal enterotoxin C2].
OBJECTIVE
To clone the gene of staphylococcal enterotoxin C2 and express it in the form of a soluble fusion protein in E. coli. Then the activation of SEC2 on mice lymphocyte and its lethal effects on tumor cells were studied.


METHODS
Staphylococcus aureus SEC2 gene was cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEC2 was used to transform E. coli BL21, where the GST-SEC2 fusion protein was expressed efficiently. The rSEC2 protein was purified with Glutathione Sepharose 4B affinity column and digested with thrombin. The in vitro culture system was utilized to observe the activation of the SEC2 on mice lymphocyte and the lethal effects on tumor cells of the activated mice lymphocyte.


RESULTS
The proper gene of SEC2 was cloned and purified rSEC2 was obtained. The MTT results indicated that rSEC2 have strong ability to stimulate mice lymphocyte to proliferate with a dose-dependent manner. With the proliferation of mice splenic lymphocyte, rSEC2 has a strong lethal effect on tumor cells B16, K562 and K562-AD.


CONCLUSIONS
In this study, the gene of SEC2 was cloned and the rSEC2 protein was obtained, which had strong lethal effect on tumor cells B16, K562 and K562-AD.
Xue
Qiao
Q
Department of Biopharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310031, China.
Ying
Yue-Bin
YB
Pan
Ying-Qiu
YQ
Li
Dan-Xi
DX
Sun
Hong-Ying
HY
Chen
Shu-Qing
SQ
D000818:Animals; D045744:Cell Line, Tumor / Q000187:drug effects; D049109:Cell Proliferation; D003001:Cloning, Molecular; D004768:Enterotoxins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics; D008213:Lymphocyte Activation / Q000187:drug effects*; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology*; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D013154:Spleen / Q000166:cytology; D014162:Transfection
D004768:Enterotoxins; D011993:Recombinant Fusion Proteins; C034959:enterotoxin C, staphylococcal; D005982:Glutathione Transferase