鸡γ-干扰素基因在大肠杆菌中的表达  

Chicken IFN-γ gene Expression in Escherichia coli

作　　者：吴树华[1] 宫鹏涛[1] 张西臣[1] 吴玲[1] 李建华[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《青岛农业大学学报（自然科学版）》2009年第1期8-11,共4页Journal of Qingdao Agricultural University(Natural Science)

基　　金：国家自然科学基金资助项目(NO.30500370);吉林省科技发展计划项目NO.20050211-2)

摘　　要：构建鸡γ-干扰素基因(lFN-γ)的融合表达质粒,并在原核系统表达。根据GenBank发表的鸡γ-干扰素核苷酸序列,使用prim er5设计一对特异性引物,通过RT-PCR技术从ConA诱导培养的鸡脾脏淋巴细胞中克隆出鸡γ-干扰素基因并对其进行测序。测序结果表明,鸡γ-干扰素基因全长495bp,具有一个完整的开放阅读框,编码164个氨基酸,与国外发表的序列同源性为100%。将序列连接到原核表达载体pET28 a(+)上,转化大肠杆菌BL21(DE3),经IPTG诱导后进行SDS-PAGE电泳分析。结果表明鸡γ-干扰素表达蛋白大小为20.8KD,并以包涵体形式表达。为鸡γ-干扰素生物制剂或疫苗佐剂的开发奠定基础。Chicken interferon -γ （IFN -γ） gene fusion expression plasmid was constructed and fusion protein expressed in prokaryotic expression system. According to the published chicken interferon -γnucleotide sequence in GenBank, a pair of specific primers were designed using the primer 5 software. The chicken interferon --γgene was amplified from the ConA - induced chicken spleen cells by RT - PCR and then sequenced. The analysis of sequence showed that the chicken interferon -γgene is 495bp in size, with an open reading frame encoding 164 amino acids. Compared with the sequence that published in GenBank,the homology is 100%. To express the chicken interferon -γ, the gene was cloned into pET - 28a （ ＋）vector, then transformed into the E. coli BL21 （ DE3 ） for expression. Induced by lmmol/L IPTG at 37℃ for 4h, the expression product was identified by SDSPAGE. The results showed that 20.8KD chicken interferon -γ protein was expressed in the form of inclusion bodies. The study laid a foundation for the development of chicken interferon -γ biological agents or vaccine adjuvant.

关 键 词：鸡γ-干扰素 克隆 原核表达 

分 类 号：S855.9[农业科学—临床兽医学]