转录因子CTCF的克隆、表达及其抗体制备  

Cloning, expression and antibody preparation of transcription factor CTCF

作　　者：何莉[1] 朱旭东[1] 毛志勇[1] 朱恒奇[1] 黄培堂[1] 

机构地区：[1]军事医学科学院生物工程研究所

出　　处：《生物技术通讯》2004年第5期454-456,共3页Letters in Biotechnology

摘　　要：用RT-PCR方法从HeLa细胞总RNA中扩增转录因子CTCF的cDNA序列,克隆至pGEM-TEasy载体上。将CTCF蛋白N段的编码序列亚克隆至pET22b(+)载体中,在大肠杆菌BL21(DE3)中诱导表达,并用镍柱进行亲和纯化。用纯化后的CTCFN段蛋白免疫家兔,获得CTCF的多克隆抗体,进而用于细胞内CTCF蛋白的免疫印迹检测。CTCF cDNA was amplified from total RNA of HeLa cells and was ligated into pGEM-T Easy plasmid. N domain of CTCF, termed CTCF-N, was expressed in E.coli BL21(DE3) using pET expression system, and then CTCF-N was purified by Ni-NTA agarose column. CTCF-N polyclonal antibody was elicited by immunizing rabbit, then purified and used to detect CTCF level by western blotting.

关 键 词：转录因子 蛋白 抗体制备 诱导表达 亚克隆 细胞内 ET 纯化 CDNA序列 总RNA 

分 类 号：Q78[生物学—分子生物学] R392[医药卫生—免疫学]