Amplification and cloning of staphylococcus aureus enterotox in D(SED) gene

  Objective   To construct a recombinant plasmid containing staphylococcus aureus toxic protein gene.

  Methods   A couple of primers were designed for PCR according to the known sequence of SED.The gene obtained by amplification from plasmid DNA of staphylococcus aureus by PCR technique was cloned into plamid of p ET32a directionally.The recombinant plasmid p ET32a was transferred into competent J M109.The recombinants were screened and identified by restriction analysis and PCR, the cloned gene was sequened.

  Results   The size of amplified PCR products was 317bp.The correct recombinant plasmid p ET32a was isolated and confirmed by restriction analysis and PCR, DNA sequencing showed the DNA sequence of the cloned gene was the same as the published sequence.

  Conclusions   The SED toxic gene was first successfully amplified and cloned into plasmid PET32a in our country.It provided the basic material for studying the pathogenesis.