鸭副粘病毒WF01 D株F基因的测序与蛋白特性分析

用鸭副粘病毒凤阳分离株WF01 D作9～10日龄SPF鸡胚的尿囊腔接种，成功增殖了该病毒，采用一步法RT-PCR技术扩增WF01 D病毒的F基因，获得了1条长约1．7kb的特异性条带。用PCR产物直接测序。测序结果表明，扩增片段大小为1782bp，含有1个1662bp的开放性阅读框，编码554个氨基酸。核苷酸同源性分析表明：WF01 D株与国内外其他NDVF基因的同源性为84．5％～97．0％，其中与国内标准强毒株F48E9的同源性为86．6％，说明WF01 D与国内外的传统毒株有较大变异。与Taiwan95株和J株的同源性为93．6％和97．0％，说明WF01 D与Taiwan95株和J株亲缘关系较近，具有较高的相似性。F蛋白裂解位点的氨基酸顺序为112Arg-Arg-Gln-Lys-Arg-Phe117，表明为NDV的强毒株。蛋白疏水性和抗原性分析表明与标准强毒F48E9株相比没有太大的变异。

Sequencing and protein analysis of F gene of WF01D from duck

The ND virus of WF01D was grown in 10-day-old SPF embrocated eggs. The F gene of Fengyang strain WF01 D of duck was amplified by one step RT-PCR. The sequencing analysis showed that the sequence of the F gene spanned 1782 bp including an ORF of 1662 bp and encoded a protein of 554 amino acids. The homology between WFol D and the NDV strain isolates inside and outside China was from 84.5% to 97.0%. The WF01 D F gene＇s nucleotide sequence shared 86.6% homology with that of the NDV strain F48E9. The ORF of WF01D F gene shared 93.6% and 97.0% nucleotide homology with that of the NDV strain Taiwan95 and J respectively. The results indicated the mutation on the F gene had varied largely,compared with classic NDV. The deduced amino acid sequences near cleavage site of F0 proteins showed a 112Arg-Arg-Gln-Lys-Arg-Phell7 motif, comforming well with that of the virulent strains. The analysis of WF01 D F protein hydrophobicity and epitope showed that, WF0, D strain was not a variation, compared with the standard strain F48E9.