[Expression and nuclease activity analysis of staphylococcus nuclease in E.coli].
OBJECTIVE
To express staphylococcus nuclease (SN) in E.coli and prepare rabbit antisera against SN.


METHODS
The SN gene was amplified by high-fidelity PCR from plasmid pPLC-SN and then subcloned into expression vector pLEX to obtain the recombinant plasmid pLEX-SN. The expression of recombinant protein was induced by tryptophan. The expressed SN was used to immunize a rabbit to prepare specific antibody.


RESULTS
SDS-PAGE analysis showed that the relative molecular mass (M(r)) of the expressed SN was about 17,000 and the expressed SN accounted for about 37% of total bacterial proteins. The prepared antisera were specific to react with recombinant SN.


CONCLUSIONS
Expression vector of SN has been successfully constructed and rabbit antibody against SN was prepared. These results lay the foundation for developing SN as antiviral protein.
Virology Lab., Institute of Microbiology and Epidemiology, AMMS, Beijing 100071, China. cfqin@hotmail.com
Cheng-feng
CF
Qin
E-de
ED
Qin
Man
M
Yu
Tao
T
Jiang
Shui-ping
SP
Chen
Yong-qiang
YQ
Deng
Hong-yuan
HY
Duan
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D007106:Immune Sera / Q000276:immunology; D008836:Micrococcal Nuclease / Q000032:analysis / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D013210:Staphylococcus / Q000201:enzymology*
D007106:Immune Sera; D008836:Micrococcal Nuclease