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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 31, No. 3, 2010, pp. 565-569
Bioline Code: zr10077
Full paper language: Chinese
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 31, No. 3, 2010, pp. 565-569

 en Expression and identification of Bm-TFF2 in Pichia pastoris
Guo-Yu, Yu; Hong-Yun, Zhang; Yong, Zhang; Ping, Jiang; Wen-Hui, Lee & Yun, Zhang

Abstract

Bm-TFF2, an amphibian trefoil factor, which is isolated from skin secretions of frog Bombina maxima, has much stronger biological activities than human TFFs. In the present study, Bm-TFF2 gene was amplified by polymerase chain reaction (PCR) from its cDNA and cloned into Pichia pastoris expression vector pPIC9K containing AOX1 promoter and α-factor leader sequence. Multi-copies insertion transformants were screened on G418 plates. After the induction by 1% methanol for 72 hours, the expression of Bm-TFF2 came up to the best quantity which was about 50 mg in 1L medium, and 80% saturation ammonium sulfate was suitable to collect the Bm-TFF2 protein, as identified by SDS-PAGE and Western blotting assay. The results showed that the plasmid of Bm-TFF2-pPIC9K was constructed successfully and expressed abundantly in eukaryotic expression system, which lies basis for researching further the biological activities and the relationship of structure and functions of Bm-TFF2.

Keywords
Bm-TFF2; Pichia pastoris; Expression; Identification

 
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