摘要
鹅副粘病毒分离株 YG97经 1 0日龄鸡胚增殖后纯化 ,提取病毒基因组 RNA,采用 RT-PCR一次性扩增出与预期设计的 1 .7kb大小相符的特异性条带。将扩增产物提纯后克隆入 p GEMR -T载体 ,经转化、筛选及酶切鉴定后 ,初步获得了含鹅副粘病毒 F基因的阳性克隆 ,并进行了序列测定。序列分析表明 :扩增的 F基因片段的长度为 1 695bp,共编码553个氨基酸 ,F蛋白裂解位点的氨基酸顺序为 1 1 2 R-R-Q-K-R-F1 1 7,与 NDV的强毒株特征相符 ,同时也与鹅副粘病毒分离株致病性试验结果相符。同源性分析表明 :与国内标准强毒株 F48E9的核苷酸同源性为 86%,与国内外部分发表的其他 NDV毒株的核苷酸同源性在 84 %～ 89%之间 ,说明该毒株相对于经典的 NDV在
The geese′s paramyxovirus isolate YG 97 was propagated in 10 day old chicken embryos. The allantonic fluids of the virus were purified and the genomic RNA was extracted. The F gene of the virus has been successfully amplified by RT PCR, and further cloned into pGEM R T vector, and a positive recombintant plasmid was screened by restriction enzyme analysis. The sequence analysis showed that the nucleotide sequence of this F gene was 1 695 bp and encoding a protein of 553 amino acids. The amino acid sequence of cleavage site region was 112 R R Q K R F 117 , matching to virulent NDV strains, and at the same time it was consistent with the pathogenicity test of the isolates. Sequence comparison with the published Chinese standard virulent strains F 48 E 9, the homology of the nucleotide was 86%, and the homology compaired with other NDV strains was between 84% and 89%. The results indicated the mutation of the F gene between the geese′s paramyxovirus and the classic NDV varied to a great extent.
