Citation: FAN Xue-zheng, WANG Qin, CHEN Zhen-hai, NING Yi-bao, WANG Zai-shi. Expression of Hog Cholera Lapinised Virus E2 Gene in E. coli and the Development of Indirect ELISA with Recombinant Protein .VIROLOGICA SINICA, 2005, 20(3) : 253-256.

Expression of Hog Cholera Lapinised Virus E2 Gene in E. coli and the Development of Indirect ELISA with Recombinant Protein

  • Available online: 20 June 2005
  • The envelope glycoprotein E2 of classical swine fever virus(CSFV) was an important protective antigen with more than thirty hydrophobic amino acids at the carboxyl terminal. E2 genes with different lengths of transmembrane region(TMR) of Hog cholera lapinised virus(HCLV) were amplified by RT-PCR,and cloned into pGEX-4T-1 to construct recombinant plasmid pGEXTE2-339, pGEXTE2-355, pGEXTE2-375, respectively. In view of codon usage in prokaryocyte, these plasmids were transfered into E.coli BL21-CodonPlus(DE3)-RP. After induction by IPTG, pGEXTE2-339, pGEXTE2-355 were successfully expressed as inclusion bodies, but pGEXTE2-375 was obviously not expressed. It was showed that TMR could decrease expression ability of HCLV E2 gene in E.coli.Furthermore,after inclusion bodies being washed and denatured, the fusion protein was used as antigen to detect antibody of CSFV and indirect ELISA was developed.

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    Expression of Hog Cholera Lapinised Virus E2 Gene in E. coli and the Development of Indirect ELISA with Recombinant Protein

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    Abstract: The envelope glycoprotein E2 of classical swine fever virus(CSFV) was an important protective antigen with more than thirty hydrophobic amino acids at the carboxyl terminal. E2 genes with different lengths of transmembrane region(TMR) of Hog cholera lapinised virus(HCLV) were amplified by RT-PCR,and cloned into pGEX-4T-1 to construct recombinant plasmid pGEXTE2-339, pGEXTE2-355, pGEXTE2-375, respectively. In view of codon usage in prokaryocyte, these plasmids were transfered into E.coli BL21-CodonPlus(DE3)-RP. After induction by IPTG, pGEXTE2-339, pGEXTE2-355 were successfully expressed as inclusion bodies, but pGEXTE2-375 was obviously not expressed. It was showed that TMR could decrease expression ability of HCLV E2 gene in E.coli.Furthermore,after inclusion bodies being washed and denatured, the fusion protein was used as antigen to detect antibody of CSFV and indirect ELISA was developed.

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