XU Fu-Zhou, CUI Chi-Zhong, LIU Yue-Long, DUAN Yu-You and SUN Fu-Chang. Construction of Physical M apping of Goose-derived Adenovirus Genomic DNA[J]. Virologica Sinica, 2000, 15(4): 388-394.
Citation: XU Fu-Zhou, CUI Chi-Zhong, LIU Yue-Long, DUAN Yu-You, SUN Fu-Chang. Construction of Physical M apping of Goose-derived Adenovirus Genomic DNA .VIROLOGICA SINICA, 2000, 15(4) : 388-394.

鹅源腺病毒基因组DNA物理图谱的构建

  • 将鹅源腺病毒Y81G4 株全基因组DNA的HindⅢ酶切片段分别插入质粒 pUC18,成功构建了全基因组DNA文库。在此基础上 ,将重组质粒携带的插入片段切出、回收并分别用地高辛标记后作为探针 ,与经限制酶BamHI、EcoRI、PstI、EcoRV消化的病毒基因组DNA进行SouthernBlotting ,杂交结果经比较综合后获得了该病毒基因组DNA的HindⅢ、BamHI、EcoRI、PstI、EcoRV限制性内切酶的物理图谱。利用已发表的含有鸡EDS76病毒AA 2株基因组DNA右末端的重组质粒pBE4 2作为探针 ,与本病毒两末端重组质粒进行SouthernBlotting ,根据同源性杂交结果确定了本病毒基因组DNA物理图谱与EDSVAA 2株相应的方向。本病毒基础因组DNA物理图谱的精确构建 ,为进行基因组结构分析 ,筛选复制非必需区 ,构建禽腺病毒载体打下了基础.

Construction of Physical M apping of Goose-derived Adenovirus Genomic DNA

  • The genomic DNA library of goose derived adenovirus (strain Y 81 G 4) was constructed after ten fragments of viral genomic DNA digested with Hin dⅢ were cloned into plasmid pUC18 respectively. The inserted fragments of the ten recombinant plasmids were labelled with digoxigenin. Viral genomic DNA digested with Bam HI\, Eco RI\, Pst I or Eco RV were transferred to NC membranes after electropheresis and probed with DIG labelled cloned Hin dⅢ fragments respectively. The physical mapping of adenovirus Y 81 G 4 genomic DNA was constructed successfully by Southern blot. To compare strain Y 81 G 4 genomic DNA mapping orientation with EDSV strain AA 2, the cloned right terminal fragment of EDSV strain AA 2 was labelled with digoxigenin and probed with the two cloned terminal fragments of Y 81 G 4 genomic DNA by Southern blot. By comparison of homology the orientation of Y 81 G 4 genomic DNA were determined. These results will be helpful for construction of avian adenovir .

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    Construction of Physical M apping of Goose-derived Adenovirus Genomic DNA

    • 1. Cot of Ani Sciand Vet Med, Yangzhou  

    Abstract: The genomic DNA library of goose derived adenovirus (strain Y 81 G 4) was constructed after ten fragments of viral genomic DNA digested with Hin dⅢ were cloned into plasmid pUC18 respectively. The inserted fragments of the ten recombinant plasmids were labelled with digoxigenin. Viral genomic DNA digested with Bam HI\, Eco RI\, Pst I or Eco RV were transferred to NC membranes after electropheresis and probed with DIG labelled cloned Hin dⅢ fragments respectively. The physical mapping of adenovirus Y 81 G 4 genomic DNA was constructed successfully by Southern blot. To compare strain Y 81 G 4 genomic DNA mapping orientation with EDSV strain AA 2, the cloned right terminal fragment of EDSV strain AA 2 was labelled with digoxigenin and probed with the two cloned terminal fragments of Y 81 G 4 genomic DNA by Southern blot. By comparison of homology the orientation of Y 81 G 4 genomic DNA were determined. These results will be helpful for construction of avian adenovir .

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