Citation: TAO Zhi-Hui, DONG Guan-Mu, SHU Yong-Xin. Cloning,Sequencing and Expressing of S Segment Coding Gene of Seoai Virus R22 Strain .VIROLOGICA SINICA, 1999, 14(4) : 322-327.

Cloning,Sequencing and Expressing of S Segment Coding Gene of Seoai Virus R22 Strain

  • Available online: 05 December 1999
  • For studing the structure and characteristic of nucleoprotein (NP) of Seoul virus R22strain, which is the seed virus of HFRS candidate vaccine, the NP gene coding region of the R22S segment was amplificated by RT-PCR and then cloned it into pET-3a expression plasmid. Se-quence assay showed that the NP gene contains 1290 nucleotides, which has homology of 96. 2%with well known Seoul virus (SR-11 ) and 71. 0% with Hantaan virus (76-118). These resultscoincide with previous Serological testings. The cloned NP gene has been transformed into E. coliBL21 and it could give a high expression when induced by IPTG. Western blot and ELISA assayof the purified products showed that expressed NP only reacted with NP-specific MAbs A35 and3D9, but not with G2-specific MA 3D8. The expressed NP can be used as a specific antigen forclinical diagnosis.

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    Cloning,Sequencing and Expressing of S Segment Coding Gene of Seoai Virus R22 Strain

    • 1. National Institute for the Control Pharmaceutical and biological 

    Abstract: For studing the structure and characteristic of nucleoprotein (NP) of Seoul virus R22strain, which is the seed virus of HFRS candidate vaccine, the NP gene coding region of the R22S segment was amplificated by RT-PCR and then cloned it into pET-3a expression plasmid. Se-quence assay showed that the NP gene contains 1290 nucleotides, which has homology of 96. 2%with well known Seoul virus (SR-11 ) and 71. 0% with Hantaan virus (76-118). These resultscoincide with previous Serological testings. The cloned NP gene has been transformed into E. coliBL21 and it could give a high expression when induced by IPTG. Western blot and ELISA assayof the purified products showed that expressed NP only reacted with NP-specific MAbs A35 and3D9, but not with G2-specific MA 3D8. The expressed NP can be used as a specific antigen forclinical diagnosis.

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