Volume 31 Issue 6
Nov.  2010
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Yu guo-yu, ZHANG Hong-Yun, ZHANG Yong, JIANG Ping, Lee Wen-Hui, Zhang Yun. Expression and identification of Bm-TFF2 in Pichia pastoris. Zoological Research, 2010, 31(6): 565-569. doi: 10.3724/SP.J.1141.2010.06565
Citation: Yu guo-yu, ZHANG Hong-Yun, ZHANG Yong, JIANG Ping, Lee Wen-Hui, Zhang Yun. Expression and identification of Bm-TFF2 in Pichia pastoris. Zoological Research, 2010, 31(6): 565-569. doi: 10.3724/SP.J.1141.2010.06565

Expression and identification of Bm-TFF2 in Pichia pastoris

doi: 10.3724/SP.J.1141.2010.06565
  • Received Date: 2010-08-17
  • Rev Recd Date: 2010-11-10
  • Publish Date: 2010-12-22
  • 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.
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