CDNA cloning of human leptin and its expression / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 476-479, 2003.
Artigo
em Chinês
| WPRIM
| ID: wpr-259164
ABSTRACT
To clone cDNA of human leptin gene and obtain leptin protein for future study on leptin binding proteins. The cDNA of human leptin with 6 x his-tag was cloned by over-hang extension PCR protocol using human genomic DNA as template, and subcloned into in vitro expression vector pIVEX2.3MCS, and the fusion protein was expressed in vitro by Rapid Translation System (RTS) (RTS500 cycle primer Kit and RTS500 ProteoMaster of Roche company). The apparent molecular weight(19.46 kD) and the immuno-specificity of the fusion protein were confirmed by SDS-PAGE and Western blot, and the expressed fusion protein stayed mainly in the supernatant of the reaction mixture in soluble form. This work provides us solid basis for further study on new leptin-associated proteins.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas Recombinantes de Fusão
/
Química
/
Western Blotting
/
Reação em Cadeia da Polimerase
/
Clonagem Molecular
/
DNA Complementar
/
Leptina
/
Eletroforese em Gel de Poliacrilamida
/
Genética
/
Metabolismo
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2003
Tipo de documento:
Artigo
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