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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (04): 17-21    DOI:
    
Cloning and Expression of the New Gene Recombinant RMBAY Against Type-2 Diabetes and Its Production Optimization
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Abstract  Constructing prokaryotic expression vector pKY-RMBAY by gene recombination and research its optimizing productive conditions. By PCR technology synthesizing the gene of the RMBAY with preference codon of E.coli and the RMBAY gene was inserted into high efficiency expression vector pKYB-MCS. Expressed fusion proteins in E.coli ER2566 were purified with Chitin-Beads column. Fusion proteins binding on Chitin-Beads was cut on N-terminus of intein due to the induction of β-mercaptoethanol and the target peptide RMBAY was released. The RMBAY was identified by mass spectrum. Experiment results showed RMBAY can be high efficiently expressed in E.coli ER2566,with optimizing productive conditions the yield of the RMBAY may be 6.7mg/L fermentation product and its purity is greater than 98%. The molecular weight of RMBAY is 3.887 kDa by mass spectrum and that accords with its theory value.

Key wordsGene recombination Production Optimization Type-2 Diabetes Mass spectrum     
Received: 01 December 2008      Published: 27 April 2009
ZTFLH:  Q785  
  Q786  
Cite this article:

MA Xi- Tu-Rong-Cha- Ceng-Le- Xie-Shan-Shan- Hong-An. Cloning and Expression of the New Gene Recombinant RMBAY Against Type-2 Diabetes and Its Production Optimization. China Biotechnology, 2009, 29(04): 17-21.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I04/17

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