Citation: LI Yue-Xi, TANG Jia-Qi, TAO Kai-Hua, JIAO Ren-Liang, LI Xian-Fu, PAN Xiu-Zhen. Cloning and Expression of NS3 Gene Fragment of HCV and IdentlficatlOil of the Purified Products .VIROLOGICA SINICA, 1994, 9(4) : 297.

Cloning and Expression of NS3 Gene Fragment of HCV and IdentlficatlOil of the Purified Products

  • Available online: 05 December 1994
  • The NS3 gene fragment of HCV was cloned from the serum of hepatitis C patients in Jiangshuprovince by RT-PCR.DNA sequencing shows the cloned NS3 cDNA contains 842bp,NcoI site and sal-I site were added respectively to the upetream and downstream primers.The cDNA was cut with NcoIand SalI,and then cloned into the NcoI and SalI sites in plasmid pBV221.The recombinant plasmidwas transformed into DH5a for gaining the E,coli expressing NS3 protein,Expressed NS3 protein(30kD)was about l7% of total proteins of the E.coli,and the purified NS3 protein by Sephacryl S-200 and IgG-Sepharose 4B chromatography(or DEAE Sepharose Fast Flow)has good antigenicity andspecificity.

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    Cloning and Expression of NS3 Gene Fragment of HCV and IdentlficatlOil of the Purified Products

    • 1. Nanjing Research Institute of Military Medicine,Nanjing 210002

    Abstract: The NS3 gene fragment of HCV was cloned from the serum of hepatitis C patients in Jiangshuprovince by RT-PCR.DNA sequencing shows the cloned NS3 cDNA contains 842bp,NcoI site and sal-I site were added respectively to the upetream and downstream primers.The cDNA was cut with NcoIand SalI,and then cloned into the NcoI and SalI sites in plasmid pBV221.The recombinant plasmidwas transformed into DH5a for gaining the E,coli expressing NS3 protein,Expressed NS3 protein(30kD)was about l7% of total proteins of the E.coli,and the purified NS3 protein by Sephacryl S-200 and IgG-Sepharose 4B chromatography(or DEAE Sepharose Fast Flow)has good antigenicity andspecificity.

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