Expression of Araneus ventricosus minor ampullate spidroin.
- Author:
Zijiang YANG
1
;
Gefei CHEN
1
;
Qing MENG
1
Author Information
1. Institute of Biological Science and Biotechnology, Donghua University, Shanghai 201620, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Escherichia coli;
genetics;
metabolism;
Fibroins;
biosynthesis;
classification;
genetics;
Genetic Vectors;
genetics;
Pichia;
genetics;
metabolism;
Recombinant Fusion Proteins;
biosynthesis;
Spiders;
classification;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2013;29(9):1323-1331
- CountryChina
- Language:Chinese
-
Abstract:
A repetitive DNA fragment, named P1, was amplified by PCR with the full-length Minor Ampullate Spidroin gene sequence of Araneus ventricosus as template. P1 was ligated with pPic3.5 and PKT expression vectors and transferred into GS115 and BL21(DE3) competence cells, respectively. SDS-PAGE and Western blot were used to analyze the recombinant his-tag fusion protein. With expressed in different expression systems, soluble P1 induced proteins could be obtained as the same size. Furthermore, the expression level and purification recovery efficiency were also higher in GS115 than that of BL21(DE3). Additionally, the expression level could be improved after optimizing the incubation and induction conditions of GS115. In this research, Pichia pastoris expression system is more suitable for the native repetitive Gly/Ala-rich spider spidroin gene sequence expression than Escherichia coli system. The data can help the native full-length MiSp gene expression and large-scale exploitation of recombinant of spider silk proteins.
