Expression of Microneme Gene mic2-7h in E. Coli

›› 1999, Vol. 1 ›› Issue (1): 67-70.

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Expression of Microneme Gene mic2-7h in E. Coli

Jiang Jianlin Jiang Jinshu

Shanghai institute of Animal Parasitology|CASSShanghai|200232 College of Animal Medicine |CAU Beijing|100094

Online:1999-06-15 Published:2009-12-28
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鸡球虫微线基因mic2-7h在大肠杆菌中的表达

蒋建林|蒋金书

中国农业科学院上海家畜寄生虫病研究所!上海200232|中国农业大学!北京100094

基金资助:
国家攀登计划85-44项目

Abstract

Abstract:

Abstract This paper report that mic2-7h gene was expressed in E. Coli. Fragment mic2-7h was excisedwith Aat Ⅱ and Not Ⅰ endonuclease from pmic2-7h, and in order to correct reading frame ,5' of mic2-7h wastreat with S1 endonuclease. Fragment of mic2-7h insert into Nhe Ⅰ site of pET28a (+),and then expressionvector pET28-mic2-7h was constructed. SDS-PAGE analysis suggested that the bacteria containing the pET-mic2-7h plasmid produce a recombinent protein of 45kD as it was induced by IPTG. Western blotting indicat-ed that reconbinant protein could react with rabbit against EtMIC-2. Property of SDS-PAGE and antigen-an-tobody reaction indicated that MIC2-7h may be a protein isoform of EtMIC-2.

Key words: Eimeria tenella Microneme mic2-7h E. Coli Gene expression

摘要：

本文在大肠杆菌中表达了鸡球虫孢子化7h微线基因mic2-7h.质粒pmic2-7h经AatⅡ酶切线化,用Sl核酸酶切平DNA分子,再用NotⅠ切出mic2-7h片段,插入表达质粒pET28-a(+)的6xHis下游NheⅠ处构建成表达载体pET28-mic2-7h.IPTG诱导表达后,经SDS-PAGE分析表明表达的重组蛋白大小为45kD,约比理论推算值(37kD)小,但Western-blotting杂交表明重组蛋白能与兔抗Etmic-2抗体反应.这进一步说明了Mic2-7h可能是Etmic-2的一种蛋白异形体.

关键词: 柔嫩艾美耳球虫, 微线基因mic2-7h, 大肠杆菌, 基因表达

CLC Number:

Q786

Jiang Jianlin Jiang Jinshu. Expression of Microneme Gene mic2-7h in E. Coli[J]. , 1999, 1(1): 67-70.

蒋建林|蒋金书 . 鸡球虫微线基因mic2-7h在大肠杆菌中的表达[J]. , 1999, 1(1): 67-70.

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Expression of Microneme Gene mic2-7h in E. Coli

鸡球虫微线基因mic2-7h在大肠杆菌中的表达

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