传染性喉气管炎病毒WG株糖蛋白gJ基因的克隆与表达  

Cloning and expression of glycoprotein J gene of infectious laryngotracheitis virus WG strain

作　　者：何来[1] 王云峰[1] 石星明[1] 崔红玉[1] 韩文雄[1] 兰德松[1] 王玫[1] 童光志[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/禽传染病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2008年第2期118-121,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家“863”计划资助项目(2003AA213020)

摘　　要：根据GenBank中已发表的传染性喉气管炎病毒全基因组序列(NC006233)设计引物,以传染性喉气管炎病毒WG株基因组为模板,PCR扩增糖蛋白gJ基因,并对其进行序列分析。用DNAStar软件分析gJ蛋白的抗原性,选择抗原性强的140aa～483aa片段,克隆并连接到原核表达载体pET-32a(+)中,转化BL21(DE3)菌株。经IPTG诱导后,获得大小为65.2ku的重组蛋白,命名为r-gJ,表达量占菌体蛋白总量的60.8%;Western blot分析表明r-gJ具有较强的抗原性。A pair of primers was designed based on the genome sequence of Infectious laryngotracheites virus （ILTV） published in GenBank （Accession no. 006233）, The glycoprotein J gene of ILTV WG strain was amplified from ILTV WG strain genome DNA, cloned into pMDIS-T vector and sequenced. Based on result of antigenicity analysis by DNAstar, a fragment of gJ, encoding 140 aa-483 aa was cloned into Prokaryotic expression plasmid pET-32a （＋） and transformed into BL21 （DE3） cells. A 65.2 ku fusion protein r-gJ was expressed in high yield and identified by SDS-PAGE after IPTG induction. Western blot showed r-gJ possessed strong antigenicity.

关 键 词：传染性喉气管炎病毒 糖蛋白J(gJ) 序列分析 表达 抗原性 

分 类 号：S852.6[农业科学—基础兽医学]