Citation: SONG Yan, SHI Dong-fang, FAN Chen, LIU Sheng-wang, LI Yi-jing, . Cloning of vp4 Gene from Porcine Rotavirus JL94 Isolate and Expression in Baculovirus System .VIROLOGICA SINICA, 2005, 20(1) : 61-64.

Cloning of vp4 Gene from Porcine Rotavirus JL94 Isolate and Expression in Baculovirus System

  • Available online: 20 January 2005
  • A pair of primers was designed to amplify vp4 gene of major antigen site (1-756 bp) of JL94 isolate. The sequence was analyzed in comparison with the VP4 amino acid sequences of two reference porcine rotavirus. The amino acid sequences were 96.43% and 67% correspondingly. Notably, the most divergence of amino acid sequence is located in a region delimited by aa81-aa207. The vp4 gene was inserted into expression plasmid pMel BacA. pMel BacA and Bac-N-blue DNA were cotransinfected insect cell Sf9. After 3 time plaque, reconstitution virus affected Sf9 and expresed in the cell. It indicated that the 30kDa product of the vp4 gene was 20% of total cell protein. Western blotting and neutralization test continmed that product had a nice biological activity. This study provides the basis for PRV identification, molecular epdiemiology investigation, and research of diagnostic reagent and genetic engineering vaccine

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    Cloning of vp4 Gene from Porcine Rotavirus JL94 Isolate and Expression in Baculovirus System

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    Abstract: A pair of primers was designed to amplify vp4 gene of major antigen site (1-756 bp) of JL94 isolate. The sequence was analyzed in comparison with the VP4 amino acid sequences of two reference porcine rotavirus. The amino acid sequences were 96.43% and 67% correspondingly. Notably, the most divergence of amino acid sequence is located in a region delimited by aa81-aa207. The vp4 gene was inserted into expression plasmid pMel BacA. pMel BacA and Bac-N-blue DNA were cotransinfected insect cell Sf9. After 3 time plaque, reconstitution virus affected Sf9 and expresed in the cell. It indicated that the 30kDa product of the vp4 gene was 20% of total cell protein. Western blotting and neutralization test continmed that product had a nice biological activity. This study provides the basis for PRV identification, molecular epdiemiology investigation, and research of diagnostic reagent and genetic engineering vaccine

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