Citation: JIANG Shi-jin, CUI Zhi-zhong, DING Jia-bo, MENG Shan-shan, YANG Han-chun. Prokaryotic Expression of pp24 Gene of Md11 Strain of Marek’s Disease Virus .VIROLOGICA SINICA, 2004, 19(4) : 369-372.

Prokaryotic Expression of pp24 Gene of Md11 Strain of Marek’s Disease Virus

  • Corresponding author: CUI Zhi-zhong, 
  • Available online: 25 August 2004
  • pp24 gene was amplified from genomic DNA of Marek’s disease viruses (MDV) Md11 strain by polymerase chain reaction(PCR) and then cloned into pGEX-6P-1 according to the right open reading frame(ORF). The recombinant plasmid was transformed into E.coli BL21 strain for expression with the induction of IPTG. SDS-PAGE and Westen-blotting with goat-anti-GST antibody validated the expression of GST-pp24 fusion protein. The expressed specific band was excised from the gel and injected into four-week-old mice 3 times, then the antiserum was collected from the mice and used for IFA with Chicken Embryonic Fibroblasts(CEF) infected by MDV Md11, CVI988 and GA strains respectively. The positive staining was found in the MDV plaques, which shows that the fusion protein of pp24 in vitro expressed has some epitopes of natural pp24.

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    Prokaryotic Expression of pp24 Gene of Md11 Strain of Marek’s Disease Virus

      Corresponding author: CUI Zhi-zhong,
    • 1. 1. Animal Science and Technology College, Shandong Agricultural University, Shandong, Taian 271018, China
    • 2. College of Veterinary Science, China Agricultural University, Beijing 100094, China

    Abstract: pp24 gene was amplified from genomic DNA of Marek’s disease viruses (MDV) Md11 strain by polymerase chain reaction(PCR) and then cloned into pGEX-6P-1 according to the right open reading frame(ORF). The recombinant plasmid was transformed into E.coli BL21 strain for expression with the induction of IPTG. SDS-PAGE and Westen-blotting with goat-anti-GST antibody validated the expression of GST-pp24 fusion protein. The expressed specific band was excised from the gel and injected into four-week-old mice 3 times, then the antiserum was collected from the mice and used for IFA with Chicken Embryonic Fibroblasts(CEF) infected by MDV Md11, CVI988 and GA strains respectively. The positive staining was found in the MDV plaques, which shows that the fusion protein of pp24 in vitro expressed has some epitopes of natural pp24.

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