伪狂犬病病毒Ea株gM和gN基因的克隆与结构分析  

Cloning and Identification of Genes Encoding for Glycoprotein M and N of Pseudorabies Virus Strain Ea

作　　者：肖少波[1] 陈焕春[1] 方六荣[1] 王革飞[1] 马相如[1] 

机构地区：[1]华中农业大学牧医学院动物病毒室,湖北武汉430070

出　　处：《中国预防兽医学报》2002年第3期171-174,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金 (39970 5 5 9) ;湖北省自然科学基金 (2 0 0 0J10 0 )

摘　　要：gM和gN是最近经经典免疫沉淀法确定的伪狂犬病病毒 (PRV)第三对异源糖蛋白二聚体。根据PRV国外Ka株的核苷酸序列 ,设计两对分别包含gM和gN完整编码区的特异性引物 ,以PRV国内地方分离株 (Ea株 )的细胞感染物为模板 ,PCR扩增出大小约 1.2kb和 0 .3kb的特异性片段。将扩增产物克隆到杆状病毒转座载体pFastBacl中 ,酶切鉴定证实后进行序列测定。结果表明 :Ea株gM、gN基因分别编码 394和 98个氨基酸 ,与Ka株的同源性分别为 98.4 %和 97.6 %。DNATool和DNASIS软件对gM、gN进行二级结构预测 ,发现gM存在 8个潜在跨膜区和一个N_糖基化位点 ,是典型的能反复多次跨膜的Ⅲ型糖蛋白 ;gN具有N_端信号肽序列、C_端跨膜区和潜在 0_糖基化位点 ,属Ⅰ型糖蛋白。上述结果为下一步深入研究gM、gN的相互作用位点及二聚体的功能奠定了基础。Glycoproteins M and N of Pseudorabies virus(PRV)can form a disulfide_linked complex.According to the nucleotide sequences of PV Ka strain,two pairs of primers corresponding to the gM and gN gene were synthesized,and the fragments containing the gM and gN coding domains of PRV Ea strain were amplified by polymerase chain reaction(PCR) from the infectious cell culture.The purified PCR products were inserted into transfer vector pFastBacl under the control of the polyhedrin promoter,resulting in plasmids pFM,pFN and the sequences were obtained by Sanger's sequencing technique.When compared with PRV Ka strain,gM and gN displayede 98.4% and 97.6% animo acid identities.The results of secondary_class analysis showed that gM containing a potencial N_glycosylation sites and eight hydrophobic domains predicted to be transmembrane sequences;gN had a hydrophobic putative signal peptide and transmembrane domains.The above results will lay the foundation for further studying the interacting sites and the funcation of the gM/gN complex.

关 键 词：伪狂犬病毒 Ea株 gM基因 gN基因 基因克隆 结构分析 

分 类 号：S855.3[农业科学—临床兽医学] Q785[农业科学—兽医学]