[Cloning and expression of human keratin 8 gene cDNA in E.coli].

AIM

To clone and express human keratin 8 gene cDNA in E.coli.

METHODS

Human cytokeratin 8 gene cDNA was amplified by RT-PCR from genomic RNA of human cell line 7721. The amplified cytokeratin 8 gene cDNA was cloned into pMD18-T vector. Then, the CK8 cDNA was amplified by PCR from recombinant plasmid pMD18-CK8, and was subcloned into pET-28a(+) expression vector. The recombinant plasmid pET-28a-CK8 DNA was transformed into E.coli DH5alpha strain.

RESULTS

Human cytokeratin 8 gene cDNA was cloned, and the recombinant plasmid pMD18-CK8 was transformed into E.coli. The CK8 cDNA was subcloned into E.coli DH5alpha strain, and successfully expressed in E.coli.

CONCLUSION

Human cytokeratin 8 gene cDNA is cloned, and successfully expressed in E.coli, which lay the foundation of further study on the CK8 biological properties and functions.