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当前位置: 首页 > 文章 > 牛环形泰勒虫裂殖子/梨浆虫表面抗原基因非疏水区的克隆及GST-P27重组蛋白的高效表达 中国畜牧兽医 2014,41 (3) 57-61
Position: Home > Articles > Clone of Tams1 Gene Non-hydrophobic Region of Theileria annulata and High Level Expression of Recombinant Protein GST-P27 China Animal Husbandry & Veterinary Medicine 2014,41 (3) 57-61

牛环形泰勒虫裂殖子/梨浆虫表面抗原基因非疏水区的克隆及GST-P27重组蛋白的高效表达

作  者：
曹雯丽;王真;沙它尔·卡哈尔;王冰洁;巴音查汗
单  位：
吐鲁番市畜牧兽医站;新疆农业大学动物医学学院
关键词：
牛环形泰勒虫;Tams1基因;克隆;GST-P27;高效表达
摘  要：
试验据GenBank登载的牛环形泰勒虫裂殖子/梨浆虫表面抗原(the merozoite/piroplasm surface antigen,Tams1)基因序列(登录号:AF214842),设计1对特异性引物,经PCR扩增出696bp的Tams1基因片段,将其插入pGEX-4T-2表达载体,提取质粒进行双酶切鉴定并测序,同源性达98%;将插入阳性质粒的BL21(DE3)菌种诱导表达重组蛋白,经优化后在37℃、0.5mmol/L IPTG诱导4h的表达条件下获得了表达量达1.39mg/mL的可溶性融合蛋白,大小为53ku;表达产物在Glutathione Sepharose 4B柱上纯化后,免疫印迹检测结果表明重组蛋白GST-P27具有良好的反应原性。
译  名：
Clone of Tams1 Gene Non-hydrophobic Region of Theileria annulata and High Level Expression of Recombinant Protein GST-P27
作  者：
CAO Wen-li;WANG Zhen;Shataer Kahaer;WANG Bing-jie;Bayinchahan;College of Veterinary Medicine,Xinjiang Agricultural University;Animal Husbandry and Veterinary Station of Turpan City;
关键词：
Theileria annulata;;Tams1gene;;clone;;GST-P27;;high level exp ression
摘  要：
A fragment about 696bp was amplified by PCR technique with specific primers based on Tams1gene sequence(AF214842)of Theileria annulata reported in GenBank.Then the target fragment was directionally cloned into pGEX-4T-2 expression vector,the recombinant plasmid DNA was cut by enzymes,and then sequenced.The result showed that the homology of the cloned Tams1gene was 98%.The positive plasmid was transformed into BL21(DE3),and then induced by IPTG. Soluble fusion protein whose expression level reached 1.39mg/mL was obtained when it was induced with 0.5mmol/L IPTG for 4hat 37℃,and it was 53ku.Western blotting showed that recombinant protein GST-P27which was purified by Glutathione Sepharose 4Bhad the favorable reactionogenicity.

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