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


CONCLUSIONS
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.
Li
Wei
W
Department of Immunology and Pathogen Biology, Medical School of Xi'an Jiaotong University, Xi'an 710061, China.
Xun
Meng
M
Chu
Yong-Lie
YL
Zheng
Jian-Wu
JW
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D053533:Keratin-8 / Q000235:genetics* / Q000378:metabolism
D018076:DNA, Complementary; C426203:KRT8 protein, human; D053533:Keratin-8