新城疫病毒(长春株)F蛋白基因的克隆和序列分析  

Cloning and Sequence Analysis of F Protein Gene of Newcastle Disease Virus Strain Changchun

作　　者：王兴龙[1,2] 金宁一[1,2] 丁壮 金扩世[1,2] 罗坤 郭志儒[1,2] 王宏伟 欧阳红生[1,2] 殷震 

机构地区：[1]解放军农牧大学军事兽医研究所 [2]白求恩医科大学生物工程研究所

出　　处：《中国兽医学报》1999年第2期109-113,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金;"863"项目

摘　　要：新城疫病毒（ＮＤＶ）长春株在鸡胚增殖后纯化，提取ＲＮＡ，然后利用特异性引物，经ＲＴ－ＰＣＲ一次性扩增出了ＮＤＶ长春株的全长Ｆ基因。将该Ｆ基因插入ｐＫＳ（－）后，进行了序列测定。序列分析表明，该Ｆ基因核苷酸长度为１７５８ｂｐ，编码５５３个氨基酸，序列中有６个糖基化位点，１３个半胱氨酸残基，裂解位点区（１１２～１１７）氨基酸序列为Ｇｌｙ－Ａｒｇ－Ｇｌｎ－Ｇｌｙ－Ａｒｇ－Ｌｅｕ，与所有弱毒株在这一区域的序列（Ｇｌｙ－Ａｒｇ／Ｌｙｓ－Ｇｌｎ－Ｇｌｙ／Ｓｅｒ－Ａｒｇ－Ｌｅｕ）相符，证明长春株为弱毒株。同源性分析表明，长春株Ｆ基因与目前国外发表的其他ＮＤＶＦ基因相比，核苷酸序列同源性在８８％～９９％之间。The NDV strain Changchun was grown in chicken embryos. The virus was purified and the genomic RNA was extracted. Using a pair of special primers, whole sequence of F gene of NDV strain Changchun was amplified by RT PCR and sequenced after being directly inserted in plasmid KS( ). The sequence analysis showed that the nucleotide sequence of this F gene was 1 758 bp long encoding a protein of 553 amino acids. In the amino acid sequence, there are 6 glycosylation sites and 13 cysteine residues. The amino acid sequence of cleavage site region (residues 112 117) is Gly Arg Gln Gly Arg Leu matching to that of all avirulent NDV strain(Gly Arg/Lys Gln Gly/Ser Arg Leu). The NDV strain Changchun was shown to be avirulent and as compared with the published NDV strains, its the homology of the nucleotide and the deduced amino acid sequence should be high.

关 键 词：新城疫病毒 F蛋白基因 RT-PCR 序列分析 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]