伪狂犬病病毒Ea株Us9基因的克隆与序列分析

对含伪狂犬病病毒Ea株BamH17片段的质粒pUC6.6进行亚克隆，构建了仅含Us9基因约0.6kb片段的重组质粒pSKMN0.6。双脱氧末端终止法进行双链测序，结果表明：Us9存在2种可能的同C-末端的编码形式，分别编码106或98个氨基酸残基，Us9基因与其下游的Us2(28K)基因之间存在约200bp的非转录区，将Ea株Us9基因序列同国外Rice株进行没源比较，发现二者在组成和结构上存在多处保守序列，尤其是潜在的酪氨酸磷酸化位点，C-端疏水性氨基酸以及由连续6个带正电荷的精氨酸残基组成的核定位信号序列，但在N-糖基化位点以及丝氨酸含量上存在较大差异。

Cloning and Sequence Analysis of the Us9 Gene of Pseudorabies Virus Ea Strain

The Recombinant plasmid pSKMN0.6,containing the complete Us9 gene of pseudorabies virus Ea strain,was constructed by subcloning from the plasmid pUC6.6,containing of the BamHI fragment 7 of Ea Strain.The whole nucleotide sequence of the 0.6kb fragment was determined by dideoxygen termination method and the result showed that there were two kinds of open reading frame,which was in_frame in C_termination and could code 106 or 98 animo acids,respectively.When compared with that of PRV Rice strain,there were multipile site_mutations in the Us9 gene of PrV Ea strain,and the diversity of amino acid residues also existed.When compared with the animo acid of Us9 of herpes simplex virus I(HSV_I),there existed several conserved founctional domains,including potential tyrosine phosphorylation site,the carboxy_terminal hydrophobic domain that function as span membrances and the string of six orginines just preceding the span membranes.On the other hand,the differences of the potential N_glycosylation sites and the percent of the serines in the whole sequences were noticed.