Amplification and cloning of shigella flexneri ipaC gene

  Objective   To constract a recombinant plasmid containing Shigella.flexneri toxic protein gene(iFaC).

  Methods   A couple of primers were designed for PCR according to the known sequence of ipaC.The ipaC gene obtained by amplification from plasmid DNA of S.flexneri by PCR technique was cloned into plasmid of pET 32a directionally.The recombinant plasmid pET 32a-ipaC was transferred into competent E.coli TG1.The recombinants were screened and identified by restriction analysis and PCR, the cloned gene was sequenced.

  Results   The size of amplified ipaC gene was 1112 bp.The correct recombinant plasmid pET 32-aipaC was isolated and confir med by restriction analysis and PCR.DNA sequencing show ed the DNA sequence of the cloned gene was the same as the published sequence.

  Conclusion   The ipaC gene was first successfully amplified and cloned into plasmid pET 32a in our country.It provided the basic material for studying the pathogenesis bacillary dysentery.