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.
Artigo
em Chinês
| WPRIM (Pacífico Ocidental)
| ID: wpr-617749
Biblioteca responsável:
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.
Texto completo:
Disponível
Base de dados:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Journal of Xi'an Jiaotong University(Medical Sciences)
Ano de publicação:
2017
Tipo de documento:
Artigo