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.
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.
出处
《中国农业科技导报》
CAS
CSCD
1999年第1期67-70,共4页
Journal of Agricultural Science and Technology
基金
国家攀登计划85-44项目
