Cloning and Prokaryotic Expression of Human Recombinant Calreticulin / 中国生物工程杂志
China Biotechnology
; (12)2006.
Artigo
em Chinês
| WPRIM (Pacífico Ocidental)
| ID: wpr-685826
Biblioteca responsável:
WPRO
ABSTRACT
Objective:
Clone, express and purify human recombinant calreticulin (CRT).Methods:
Human CRT cDNA was amplified from total RNA of human lung cancer cell line A549 cells by RT-PCR. Then, PCR product was subcloned into prokaryotic expression vector pET-15b. After sequencing, this recombinant plasmid was transformed into E.coli. Rossetta. Recombinant CRT was expressed in host cells by IPTG induction. Resulted protein was purified by Ni-NTA resin under denature condition and dialyzed to recover its native structure. SDS-PAGE and Western blot method were used to identify the expression and purification of reconbinant CRT.Results:
Human CRT cDNA was cloned from total RNA of A549 cells. CRT prokaryotic expression vector pET-15b-crt was constructed. Reconbinant CRT was induced to express in E.coli and purified by Ni-NTA affinity chromatograph.Conclusion:
A method for prokaryotic expression and purification of human recombinant CRT was successfully established. This method laid a foundation for the subsequent CRT research.
Texto completo:
Disponível
Base de dados:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
China Biotechnology
Ano de publicação:
2006
Tipo de documento:
Artigo