Research on secretion expression in Pichia pastoris and function of the HC-pro gene of watermelon mosaic virus / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1102-1106, 2007.
Article
em Zh
| WPRIM
| ID: wpr-276154
Biblioteca responsável:
WPRO
ABSTRACT
HC-pro gene of Watermelon Mosaic virus was obtained by RT-PCR was 1371bp in length. It was cloned into pPI(9K, then the eucaryotic recombinant expression plasmid pPIC9K-WHC was constructed. After being linearized with restriction endonuclease Sal I , the recombinant plasmid was transformed into Pichia pastoris GS115 by electroporation. The high copy transformants with Mut+ /His+ phenotype were selected by RT-PCR and screening on G418, MD and MM medium. Induced by methanol for 5 days, the culture supernatant was analyzed by SDS-PAGE, the results showed that a specific protein with a molecular weight of about 66 kD was expressed. Western blot analysis proved that the expression protein could specifically bind to HC-Pro polyclonal antibody. Far western blot analysis proved that the expression protein could bind to coat protein, given support to "bridge" hypothesis that HC-Pro help aphid transmission of non-persistent viruses.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Pichia
/
Proteínas Virais
/
Virologia
/
Proteínas Recombinantes
/
Cisteína Endopeptidases
/
Citrullus
/
Genética
/
Metabolismo
/
Vírus do Mosaico
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2007
Tipo de documento:
Article