High-level expression of fusion protein GGH in Pichia pastoris GS115 by constructing a double plasmid co-expression system / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 983-989, 2011.
Article
Dans Chinois
| WPRIM
| ID: wpr-324512
ABSTRACT
In order to make a large-scale preparation of(GLP-1A2G)2-HAS (GGH), the double-plamid pPICZalphaB and pPIC9K co-expression system was introduced into Pichia pastoris GS115. Firstly, the GGH fusion gene was amplified by PCR to create the recombinant expression plasmid pPICZalphaB-ggh, which was transformed into P. pastoris GS 115/F2 that was integrated by another recombinant expression plasmid pPIC9K-ggh. The immunology method combined with high concentration antibiotic was used to screen recombinant strain P. pastoris GS115/F3 capable of high-level expression of GGH protein. The GGH fusion protein expressed by GS115/F3 increased 49.7% compared with the GS 115/F2 in the expression conditions of 3% methanol inducing 80 h at 30 degrees C. At the same time, the quantitative PCR results showed that GGH gene dose in GS115/F3 increased 26.7% with respect to that of GS 115/F2. Furthermore, the Western blotting experiment indicated that the recombinant GGH possess the two antigenicities of GLP-1 and HSA.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Pichia
/
Plasmides
/
Protéines de fusion recombinantes
/
Sérumalbumine
/
Réaction de polymérisation en chaîne
/
Glucagon-like peptide 1
/
Vecteurs génétiques
/
Génétique
/
Métabolisme
/
Méthodes
Limites du sujet:
Humains
langue:
Chinois
Texte intégral:
Chinese Journal of Biotechnology
Année:
2011
Type:
Article
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