Prokaryotic expression and purification of the cDNA of recombinant human cytokeratin 9 / 西安交通大学学报(医学版)
Journal of Xi'an Jiaotong University(Medical Sciences)
; (6): 502-506, 2017.
Article
in Zh
| WPRIM
| ID: wpr-617749
Responsible library:
WPRO
ABSTRACT
Objective To clone and fuse the cDNA of human cytokeratin 9 in prokaryotic expression system,and purify and identify the fusion protein.Methods The cDNA fragment of human cytokeratin 9 was amplified from human keratinocyte (HaCaT) total RNA with specific primers.The PCR products were cloned into vector pET-28a,then the fusion protein of his-CK9 was induced by IPTG.The expressed fusion protein of his-CK9 was purified by nickel ion affinity chromatography and identified by SDS-PAGE and Western blot.Results The sequencing proved that the recombinant vector of the cDNA of CK9 was correct.The fusion protein of his-CK9 was induced to be expressed in E.coli.The fusion protein of his-CK9 was highly purified and his-CK9 showed specific binding to the commercialized antibodies of CK9.Conclusion The recombinant vector of pET-28a-CK9 has been successfully constructed,and the fusion protein of his-CK9 has been successfully expressed and purified.
Full text:
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Database:
WPRIM
Language:
Zh
Journal:
Journal of Xi'an Jiaotong University(Medical Sciences)
Year:
2017
Document type:
Article