鹅细小病毒长春分离株主要结构蛋白(VP2-VP3)基因的原核表达  

Expression of the major structural protein(VP2-VP3)gene of a Chinese isolate,goose parvovirus in E.coli

作　　者：李雪梅[1,2] 章金刚[1,3] 向华[1] 陈晓月[1] 金宁一[4] 费恩阁[1] 

机构地区：[1]解放军军需大学动物科技系 [2]清华大学结构生物学实验室,北京100084 [3]军事医学科学院输血医学研究所,北京100850 [4]解放军军需大学军事兽医研究所

出　　处：《生物技术通讯》2002年第5期331-333,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(39870557);教育部和总后勤部回国人员启动基金项目(98H026)

摘　　要：根据国外已发表的鹅细小病毒(GPV)A株基因组核苷酸序列,设计并合成了一对用于GPV主要结构蛋白(VP2-VP3)基因表达的引物。利用合成的引物,扩增并鉴定了GPV中国长春株(GPVCC株)的VP2-VP3基因。将扩增的目的基因插入到原核表达载体pET28(a)的多克隆位点,构建了表达GPVVP2-VP3基因的原核载体pEGVP1。重组表达载体质粒转化BL21宿主菌,经IPTG诱导,SDS-PAGE检测到大小分别为75000和58000的表达蛋白带。Westernblot分析表明,表达产物具有很好的特异性。According to nucleotide suquence of GPV A strain,a pairs of primers were designed and synthesized,and mutated the VP2ACG codon to ATG one.The complete VP2-VP3(the major structural protein)genes of a Chinese isolate stain(GPV CC)was amplified by PCR,and obtained a DNA fragment with the sizes of1.8kb.The PCR prod-uct was digested with Bam HI and the resulting fragment was inserted into the Bam HI site of the pET28(a)under the downstream sequences.E.coli component host BL21was transformed with the recombinant.The recombinant bac-teria could express the VP2-VP3gene after induced by IPTG,and the recombinant protein were analyzed on12%SDS-PAGE and stained with comass brilliant blue or had been detected by immunblotting analysis with anti-GPV serum.

关 键 词：鹅细小病毒 长春分离株 结构蛋白 (VP2-VP3) 原核表达 基因表达 

分 类 号：S852.659.2[农业科学—基础兽医学]