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Volume 21 Issue 1
February 2006
    Citation: LI Dou-lin, LI Yan-qiu, ZHANG Jia-min, YANG Bo, CHENG Wu-guo, ZHOU Wei-dong, HU Yuan-yang*. Expression and Localization of the Dendrolimus punctatus CypovirusPolyhedrin gene .VIROLOGICA SINICA, 2006, 21(1) : 52-56.
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    Expression and Localization of the Dendrolimus punctatus CypovirusPolyhedrin gene

    • 1.

      State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China

    • Available online: 20 January 2006
    • To express the polyhedrin (the tenth segment, S10) of Dendrolimus punctatus Cypovirus (DpCPV) and investigate the localization of the polyhedrin in eukaryote, The polyhedrin gene was amplified from DpCPV by RT-PCR and cloned into pET-28a vector. The polyhedrin was then expressed in bacteria. The purified protein was used to immunized rabbit to produce the antibody. The S10 ORF was also cloned into the baculovirus vector pFASTBAC-HTb. The recombinantplasmid pFASTBACS10 was transformed to E. coli containing AcMNPV bacmid to generate the recombinant Bacmid AcS10. AcS10 bacmid DNA transefect Sf9 cell and the recombinant baculoviruse vAcS10 was produced. After S10 was expressed, the products were detected and analyzed by the SDS-PAGE, Western-blot and Immunofluorescence technique. The polyhedrin expressed in SF-9 cells was mainly located in cytoplasm.
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      Expression and Localization of the Dendrolimus punctatus CypovirusPolyhedrin gene

      • 1. State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China

      Abstract: To express the polyhedrin (the tenth segment, S10) of Dendrolimus punctatus Cypovirus (DpCPV) and investigate the localization of the polyhedrin in eukaryote, The polyhedrin gene was amplified from DpCPV by RT-PCR and cloned into pET-28a vector. The polyhedrin was then expressed in bacteria. The purified protein was used to immunized rabbit to produce the antibody. The S10 ORF was also cloned into the baculovirus vector pFASTBAC-HTb. The recombinantplasmid pFASTBACS10 was transformed to E. coli containing AcMNPV bacmid to generate the recombinant Bacmid AcS10. AcS10 bacmid DNA transefect Sf9 cell and the recombinant baculoviruse vAcS10 was produced. After S10 was expressed, the products were detected and analyzed by the SDS-PAGE, Western-blot and Immunofluorescence technique. The polyhedrin expressed in SF-9 cells was mainly located in cytoplasm.

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