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Volume 13 Issue 4
August 1998
    Citation: BI Lan, LEI Xiao-Jun, CHEN Wei, YAN Jia-Xin, YU Mo-Song, SHU Jia-Hong. High-level expression of HIV-1 gp41 gene and purification of its product .VIROLOGICA SINICA, 1998, 13(4) : 344.
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    High-level expression of HIV-1 gp41 gene and purification of its product

    • 1.

      Department of Engineering．Wuhan lnstitute of Biological ProductsMinistry of Public Health，Wuhan 430O6O

    • Available online: 05 December 1998
    • In order to get high level expression of HIV 1 gp41 gene, a fragment (690 bp) encoding N terminal of gp41 from all sequence of HIV 1 was amplified by PCR. After digesting by restriction enzyme EcoRI/Sal I, the fragment was cloned into pET28a plasmid which was digested by the same restriction enzyme, then recombinant plasmid was transformed into the E.coli BL21 (DE3). After inducing by IPTG, the bacteria produced the protein. Then the protein was purified through chelating Sepharose. Using indirect ELISA, SDS PAGE and Western blot test, it was found that the protein had good antigenicity, good specificity and high purification (99%), and its yield was about 45% of the total bacteria protein.
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      High-level expression of HIV-1 gp41 gene and purification of its product

      • 1. Department of Engineering．Wuhan lnstitute of Biological ProductsMinistry of Public Health，Wuhan 430O6O

      Abstract: In order to get high level expression of HIV 1 gp41 gene, a fragment (690 bp) encoding N terminal of gp41 from all sequence of HIV 1 was amplified by PCR. After digesting by restriction enzyme EcoRI/Sal I, the fragment was cloned into pET28a plasmid which was digested by the same restriction enzyme, then recombinant plasmid was transformed into the E.coli BL21 (DE3). After inducing by IPTG, the bacteria produced the protein. Then the protein was purified through chelating Sepharose. Using indirect ELISA, SDS PAGE and Western blot test, it was found that the protein had good antigenicity, good specificity and high purification (99%), and its yield was about 45% of the total bacteria protein.

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