Citation: Xue-Xiao-Beng, XU Zhi-Kai, MA Wen-Yu, YAN Yan, YIN Wen, ZHANG Fang-Lin, TUN Xin-An, DIAO Qian, BAI Wen-Chao. Cloning,Sequencing and Expression of S Gene Encoding Region 0f Han taan Virus Strain Chen .VIROLOGICA SINICA, 1999, 14(1) : 48-51.

Cloning,Sequencing and Expression of S Gene Encoding Region 0f Han taan Virus Strain Chen

  • Available online: 05 March 1999
  • RNA of Hantaan virus (HNTV) strain Chen isolated from China was extracted from lysate of Vero E 6 cell infected by the virus. With the RNA as template, 1.3kb cDNA fragment containing the region encoding nucleocapsid protein (NP) was obtained by reverse transcription polymerase chain reaction (RT PCR). This fragment was cloned into pGEM 7zf(+) plasmid and sequenced. Homology comparision showed that the homology of the nucleotide and amino acid sequences between strain Chen and strain 76 118 was 86% and 97%, respectively. The RT PCR product was cloned into pGEX 4T 1 vector and was efficiently expressed in E.coli . The expressed product is NP GST fusion protein with molecular weight about 70 kD in SDS pAGE. Its antigenic epitope was recognized by Western blotting and ELISA using the McAbs. The results showed that there were some differences on antigenic epitopes between strain Chen and strain 76 118.

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    Cloning,Sequencing and Expression of S Gene Encoding Region 0f Han taan Virus Strain Chen

    • 1. Dept Microhldogy,The Fourth Military Medical Uniersity,Xi'an 710032

    Abstract: RNA of Hantaan virus (HNTV) strain Chen isolated from China was extracted from lysate of Vero E 6 cell infected by the virus. With the RNA as template, 1.3kb cDNA fragment containing the region encoding nucleocapsid protein (NP) was obtained by reverse transcription polymerase chain reaction (RT PCR). This fragment was cloned into pGEM 7zf(+) plasmid and sequenced. Homology comparision showed that the homology of the nucleotide and amino acid sequences between strain Chen and strain 76 118 was 86% and 97%, respectively. The RT PCR product was cloned into pGEX 4T 1 vector and was efficiently expressed in E.coli . The expressed product is NP GST fusion protein with molecular weight about 70 kD in SDS pAGE. Its antigenic epitope was recognized by Western blotting and ELISA using the McAbs. The results showed that there were some differences on antigenic epitopes between strain Chen and strain 76 118.

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