[Expression and nuclease activity analysis of staphylococcus nuclease in E.coli]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 Jul;21(4):513-5.
[Article in Chinese]

Abstract

Aim: 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.

Conclusion: 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.

MeSH terms

  • Animals
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics*
  • Gene Expression
  • Genetic Vectors / genetics
  • Immune Sera / immunology
  • Micrococcal Nuclease / analysis
  • Micrococcal Nuclease / genetics*
  • Micrococcal Nuclease / immunology
  • Micrococcal Nuclease / metabolism*
  • Polymerase Chain Reaction
  • Staphylococcus / enzymology*

Substances

  • Immune Sera
  • Micrococcal Nuclease