Citation: CHEN Wei-Min, CUI Chi-Zhong, DUAN Yu-You, LIU Yue-Long. Amplification and Cloning of Chicken Anemia Virus whole genomic DNA by Polymerase Chain Reaction .VIROLOGICA SINICA, 1999, 14(1) : 80-86.

Amplification and Cloning of Chicken Anemia Virus whole genomic DNA by Polymerase Chain Reaction

  • Available online: 05 March 1999
  • By two pairs of primers complementory to the published sequences at the both sides of EcoRI site and BamHI site on CAV circular genome, two DNA fragments containing two parts (1.5 kb and 0.8 kb) of CAV genome divided by EcoRI and BamHI sites were amplified by PCR from the circular CAV genome (2.3 kb) detected by dot blot in genomic DNA of CAV infected RP1 cells. Through re ligation of the selected sequences from the amplified fragments, the whole CAV genomic DNA was cloned into pUC18, and the recombinant plasmid was designated as pCAV2.4. Restriction endonucleases analysis of pCAV2.4 showed that there were sites of BamHI、PstI、HindⅢ,but no site of EcoRI. Two ends of cloned DNA sequence analysis showed that the recombinant plasmid pCAV2.4 containes the whole genomic DNA sequence of CAV, and that the site sequence of EcoRI in the cloned DNA was changed by one base substitution.

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    Amplification and Cloning of Chicken Anemia Virus whole genomic DNA by Polymerase Chain Reaction

    • 1. College Animal Science and Veterinary Medicine Yangzhou University,Yangzhou 225009

    Abstract: By two pairs of primers complementory to the published sequences at the both sides of EcoRI site and BamHI site on CAV circular genome, two DNA fragments containing two parts (1.5 kb and 0.8 kb) of CAV genome divided by EcoRI and BamHI sites were amplified by PCR from the circular CAV genome (2.3 kb) detected by dot blot in genomic DNA of CAV infected RP1 cells. Through re ligation of the selected sequences from the amplified fragments, the whole CAV genomic DNA was cloned into pUC18, and the recombinant plasmid was designated as pCAV2.4. Restriction endonucleases analysis of pCAV2.4 showed that there were sites of BamHI、PstI、HindⅢ,but no site of EcoRI. Two ends of cloned DNA sequence analysis showed that the recombinant plasmid pCAV2.4 containes the whole genomic DNA sequence of CAV, and that the site sequence of EcoRI in the cloned DNA was changed by one base substitution.

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