H3N8亚型马流感病毒HA1基因原核表达载体的构建及表达  

Construction of Prokaryotic Expression Vector of H3N8 Subtype Equine Influenza Virus HA1Gene of and Its Expression

作　　者：何晶[1] 马伟[1] 孙建华[1] 徐泉源[1] 冉多良[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《新疆农业大学学报》2013年第1期12-15,共4页Journal of Xinjiang Agricultural University

基　　金：科技部-国家星火计划重大专项(2011GA890008);新疆维吾尔自治区重大专项(201130101-1-6)

摘　　要：采用RT-PCR方法从马流感病毒中国分离株A/马/新疆/07(H3N8)中扩增的HA1基因片段,将其克隆到表达载体pET-28a(+)中,测序正确后转化入Rosetta(DE3)中进行IPTG诱导表达。结果表明,重组菌表达出约53ku,以沉淀性形式存在的重组蛋白,且在37℃,0.4mmol/L IPTG条件下诱导8h表达效果最好。表达产物经切胶纯化后得到纯度较高的目的蛋白。经Western-blot分析,表达的蛋白与抗马流感病毒阳性血清发生特异性反应,证实该蛋白具有良好的反应原性。The HA1 gene was amplified from equine influenza virus（EIV）A/equine/Xinjiang/07（H3N8） strain by RT-PCR and cloned into the pET-28a（＋） expression vector. After sequencing, the recombinant plasmid was transformed into Rosetta （DE3） and expressed IPTG induction. The results indicated that the recombinant bacterial produced 53 ku mostly,existed in recombinant protein in soluble form and induced for 8 h under the conditions of 37 ~C ,0.4 mmol/L IPTG with the best effects. The purer objective protein was obtained after expression products were purified. The Western-blot was analyzed, there was peculiar reaction between expressed protein and the positive serum of resistant equine influenza virus. These results indicated that the protein had good antigencity.

关 键 词：H3N8亚型马流感病毒 HA1基因 克隆 原核表达 

分 类 号：S851.659.5[农业科学—预防兽医学]