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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (06): 1-06    DOI:
    
Expression of Gene Recombinant GLP-1 Derived Polypeptide and Its Purification and Identification
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Abstract  

Abstract In order to study the optimal expression and purification condition of the GLP-1 derived polypeptide, by PCR technology synthetizing the gene of the GLP-1 derived polypeptide with preference codon of E.coli with the plasmid containing human wild-type GLP-1 gene. Expressed fusion proteins were purified and desalted with Ni-NTA column and C18 Sep-Pak column, respectively. After chemical cleavaged by formic acid hydroformicant, the hydrolysis products were purified with Ni-NTA column and HPLC. The target peptide was identified by mass spectrum. Experiment results showed in E.coli BL21 the optimal expression condition as follow:inducing temperature is 37℃, inducing time is 6h, and the concentration of the IPTG is 0.6mmol/L. The optimal chromatographic condition of getting HPLC as follow:mobile phase A (10% CNCH3∶90% H2O,0.1%TFA),mobile phase B(100% CNCH3,0.1% TFA),flow rate is 1ml/min, 30 min of the linear gradient elution, B phase reaches 70% and the detection wave length is 280nm. The molecular mass of the GLP-1 derived polypeptide is 5.492 kDa,through mass spectrum identification. The yield of GLP-1 derived polypeptide prepared with the optimal expression and purification condition may reach 11.6mg/L fermentation product and its purity is equal or greater than 98%.



Received: 21 October 2008      Published: 02 July 2009
ZTFLH:  null  
Corresponding Authors: MA Yi     E-mail: tlihj@163.com
Cite this article:

MA Xi- Li-Hong-Jian- Zhou-Tian-Hong. Expression of Gene Recombinant GLP-1 Derived Polypeptide and Its Purification and Identification. China Biotechnology, 2009, 29(06): 1-06.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I06/1

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