【摘要】 利用 RT PCR 技术扩增出了 4 株 NDV 分离株(HeB 4 99, HeB 5 99、HeB 6 02 和HLJ 10 02)F基因539 bp的片段,将该片段克隆到 pMD 18 T载体上。经酶切分析、质粒 PCR鉴定及核苷酸序列测定,成功获得了4株 NDV分离株 F基因部分片段的重组质粒。同源性分析显示,4株分离毒株的核苷酸序列具有 95.9%~100%的同源性,与 LaSota的核苷酸序列同源性为81.0%~81.8%,与F48E9的核苷酸序列同源性达85.8%~89.3%。推导氨基酸序列分析表明,4个毒株F蛋白的裂解位点氨基酸组成为112RRQKRF117,具有强毒株裂解位点氨基酸组成特点,与ICPI和MDT测定结果相吻合。73株NDV毒株的系统发育进化树分析表明,HeB 4 99、HeB 5 99、HeB 6 02和HLJ 10 02均归属于基因Ⅶ型。更多还原

【Abstract】 With a pair of primers designed and synthesized, the 539bp fragments were amplified from four NDV strains from Heilongjiang and Hebei provinces by RT-PCR and then cloned into the pMD18-T vector. The recombinant plasmid was proved to be positive by enzyme analysis and plasmid PCR. The （nucleotide） sequence analysis showed that the four strains shared from （95.9%）to 100% homology among themselves, from 81.0% to 81.8% with NDV LaSota, and from 85.8% to 89.3% withNDV F48E9 strain, respectively. The deduced amino acids showed that special sequence of F cleavage site that had the same amino acids （pattern） as velogenic strain was (¹¹²RRQKRF¹¹⁷,) which was accordant with （the） results of ICPI and （MDT.） On the basis of the first （374bp） nucleotides initiated from the first ATG of F （gene,） a phylogenetic tree of （73） NDV strains including the four strains, the other strains isolated recently in China and the some strains （abroad） was created. The phylogenetic analysis showed that the four strains belonged to Group Ⅶ, which was a novel genotype of NDV in China.更多还原