鸡白细胞介素2基因的克隆、原核表达及功能研究

CORC > 集美大学

	鸡白细胞介素2基因的克隆、原核表达及功能研究
	郭川 ; 郑忠辉 ; 吴国平 ; 林由 ; 苏文金 ; 曹敏杰
刊名	http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=fzdz200605029&dbcode=CJFQ&dbname=CJFQ2006
	2012-06-05 ; 2012-06-05
关键词	鸡白细胞介素2 克隆原核表达免疫印迹 chicken interleukin-2 cloning expression Western blot S852.4
其他题名	Study on the cloning,expression and function of chicken interleukin-2
中文摘要	以RT-PCR法从鸡脾脏细胞中扩增出鸡白细胞介素2编码基因,通过双酶切、连接步骤克隆进原核表达载体pQE30及pGEX-4T-3,构建了鸡白细胞介素2基因的重组原核表达质粒pQE30-chIL-2及pGEX-chIL-2.重组质粒转化大肠杆菌JM109后经诱导表达,分别得到两种表达产物.以SDS-PAGE和Westernblot检测确认表达产物为带6组氨酸标签及GST蛋白的融合鸡白细胞介素2,分子量分别为16 kDa及39.5 kDa.表达蛋白与抗鸡白细胞介素2单抗均有良好的免疫结合性,进一步证实为鸡白细胞介素2.表达产物经过纯化复性后,具有明显促进鸡脾淋巴细胞增殖的作用.; The gene encoding chicken interleukin-2(chIL-2) was amplified from chicken spleen cells by RT-PCR.Following double digestion by restriction endonucleases and ligation,chIL-2 was cloned into prokaryotic expression vectors of pQE30 and pGEX-4T-3 to construct recombinants of pQE30-chIL-2 and pGEX-chIL-2,which were consequently transformed into E.coli JM109.Two kinds of recombinant protein were obtained with IPTG induction.The molecular weights of the expressed proteins were 16?kDa and 39.5?kDa on SDS-PAGE and they were regarded as 6His and GST labeled protein(respectively) as checked by Western blot.Both proteins revealed specific reaction to anti-chIL-2 mAb,further confirming that recombinant proteins are chicken interleukin-2.After purification and(renaturation),the 6His-chIL-2 showed biological activity in stimulating chicken lymphocyte proliferation.; 【作者单位】集美大学生物工程学院；厦门大学生命科学学院；集美大学生物工程学院福建厦门361021；福建厦门361005；集美大学水产学院；福建厦门361021；【作者英文名】GUO Chuan~(1,2),ZHENG Zhong-hui~2,WU Guo-ping~(2,3),LIN You~1,SU Wen-jin~1,CAO Min-jie~1(1.School of Biotechnology,Jimei University,Xiamen,Fujian 361021,China;2.School of Life Science,Xiamen University,Xiamen,Fujian 361005,China;3.School of Fisheries
语种	中文
内容类型	期刊论文
源URL	[http://ir.calis.edu.cn/hdl/235041/18513]
专题	集美大学
推荐引用方式 GB/T 7714	郭川,郑忠辉,吴国平,等. 鸡白细胞介素2基因的克隆、原核表达及功能研究[J]. http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=fzdz200605029&dbcode=CJFQ&dbname=CJFQ2006,2012, 2012.
APA	郭川,郑忠辉,吴国平,林由,苏文金,&曹敏杰.(2012).鸡白细胞介素2基因的克隆、原核表达及功能研究.http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=fzdz200605029&dbcode=CJFQ&dbname=CJFQ2006.
MLA	郭川,et al."鸡白细胞介素2基因的克隆、原核表达及功能研究".http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=fzdz200605029&dbcode=CJFQ&dbname=CJFQ2006 (2012).