Volume 32 Issue 4
Jul.  2011
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yu guo-Yu, XIANG Yang, ZHANG Hong-Yun, JIANG Ping, Lee Wen-Hui, ZHANG Yun, ZHANG. Expression of Bm-TFF2 mutants in Escherichia coli andtheir cell migration-promoting activity. Zoological Research, 2011, 32(4): 379-385. doi: 10.3724/SP.J.1141.2011.04379
Citation: yu guo-Yu, XIANG Yang, ZHANG Hong-Yun, JIANG Ping, Lee Wen-Hui, ZHANG Yun, ZHANG. Expression of Bm-TFF2 mutants in Escherichia coli andtheir cell migration-promoting activity. Zoological Research, 2011, 32(4): 379-385. doi: 10.3724/SP.J.1141.2011.04379

Expression of Bm-TFF2 mutants in Escherichia coli andtheir cell migration-promoting activity

doi: 10.3724/SP.J.1141.2011.04379
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  • Author Bio:

    yu guo-Yu,XIANG Yang

    yu guo-Yu,XIANG Yang

  • Corresponding author: ZHANG Yong
  • Received Date: 2011-01-05
  • Rev Recd Date: 2011-05-18
  • Publish Date: 2011-08-22
  • Bm-TFF2, a trefoil factor from the large-webbed bell toad (Bombina maxima), can stimulate cell migration and inhibit cell apoptosis. To study the structure-function relationship of Bm-TFF2, we constructed wild-type and mutated Bm-TFF2 plasmids and expressed recombinant proteins in E. coli. The wild-type Bm-TFF2 gene encoding mature peptide was obtained by RT-PCR, while the N-terminal, C-terminal and two arginine mutated Bm-TFF2 clones were constructed, and ligated into pET-32a(+) expression vectors. The fusion proteins were induced by IPTG at 37°C. The mutant Bm-TFF2 fusion proteins expressed mainly in the inclusion bodies. The mutant (TRX)/Bm-TFF2 could be purified by using Ni2+-chelating chromatography and reverse-phase HPLC from the inclusion body supernatant. The fusion proteins were analyzed by SDS-PAGE and Western blotting. The yield of mutant Bm-TFF2 fusion proteins of above 95% purity was about 20 mg/L. All three recombinant mutant proteins can promote the migration of AGS cells in a dose-dependent manner with no obvious activity difference.
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