| 抗隐孢子虫子孢子ScFv-PE40免疫毒素表达质粒的构建及其在大肠杆菌中的表达 |
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| 引用本文： | 尹继刚,张西臣,李建华,朱平,柳增善. 抗隐孢子虫子孢子ScFv-PE40免疫毒素表达质粒的构建及其在大肠杆菌中的表达[J]. 寄生虫与医学昆虫学报, 2005, 12(3): 139-142 |
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| 作者姓名： | 尹继刚 张西臣 李建华 朱平 柳增善 |
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| 作者单位： | 吉林大学畜牧兽医学院,长春,130062;吉林大学畜牧兽医学院,长春,130062;吉林大学畜牧兽医学院,长春,130062;吉林大学畜牧兽医学院,长春,130062;吉林大学畜牧兽医学院,长春,130062 |
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| 基金项目： | 国家自然科学基金资助项目(编号39970563) |
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| 摘 要： | 用PCR方法扩增抗隐孢子虫子孢子ScFv片段,插入表达载体pET-28a中,构建重组质粒pET-28-ScFv,然后,将绿脓杆菌外毒素(PE40)片段定向克隆到ScFv基因的下游,构建免疫毒素表达质粒pET28ScFv-PE40。经酶切分析、PCR检测和测序进行鉴定。成功地构建了免疫毒素表达质粒pET28ScFv-PE40。将上述质粒转化受体菌BL21(DE3)后,经IPTG诱导,成功地表达了目的蛋白。免疫毒素ScFv-PE40大小约为66kDa,表达量约占菌体蛋白总量的14%。
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| 关 键 词： | 隐孢子虫 子孢子 ScFv-PE40 免疫毒素 |
| 修稿时间： | 2004-10-11 |
CONSTRUCTION OF ANTI-CRYPTOSPORIDIUM PARVUM SPOROZOITE IMMUNOTOXIN ScFv-PE40 AND ITS EXPRESSION IN E. COLI |
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| YIN Ji-Gang,ZHANG Xi-Chen,LI Jian-Hua,ZHU Ping,LIU Zeng-Shan. CONSTRUCTION OF ANTI-CRYPTOSPORIDIUM PARVUM SPOROZOITE IMMUNOTOXIN ScFv-PE40 AND ITS EXPRESSION IN E. COLI[J]. Acta Parasitologica et Medica Entomologica Sinica, 2005, 12(3): 139-142 |
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| Authors: | YIN Ji-Gang ZHANG Xi-Chen LI Jian-Hua ZHU Ping LIU Zeng-Shan |
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| Abstract: | ScFv fragments were amplified by PCR,and digested with restriction endonuclease,then inserted into expressional plasmid,pET 28a to construct a recombinant plasmid pET28-ScFv.Fragments of pseudomonas aeruginosa exotoxin(PE40)were inserted into the downstream site of ScFv on the expressional plasmid,pET-28-ScFv.The constructed plasmid pET28-ScFv-PE40 was identified by enzyme digestion analysis,PCR and sequencing.After transforming these plasmids into E.coli strain BL21(DE3)and induced by IPTG,immunotoxin ScFv-PE40 was proved to be expressed successfully.The expressed recombinant immunotoxins ScFv-PE40 takes 14% of the total cell proteins. |
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| Keywords: | Cryptosporidium parvum Sporozoite ScFv-PE40 Immunotoxin |
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