人小肠三叶因子的克隆及酶联免疫吸附检测

北京大学学报（自然科学版）

人小肠三叶因子的克隆及酶联免疫吸附检测

口如琴,王蔚,李令媛,茹炳根

北京大学生命科学学院，生物化学及分子生物学系，蛋白质工程国家重点实验室，北京，100871

收稿日期:1999-09-07 出版日期:2000-11-20 发布日期:2000-11-20

Clone and Enzyme-Linked Immunosorbent Assay of Human Intestinal Trefoil Factor

KOU Ruqin，WANG Wei，LI Lingyuan，RU Binggen

Department of Biochemistry and Molecular Biology, College of Life Sciences, Peking University, National Laboratory of Protein Engineering, Beijing, 100871

Received:1999-09-07 Online:2000-11-20 Published:2000-11-20

摘要/Abstract

摘要： 人小肠三叶因子的cDNA，克隆到原核表达载体pGEX-4T-1中，经IPTG诱导后，hITF以GST-hITF融合蛋白的形式表达，融合蛋白占细菌总蛋白的15%左右。经过Glutathione-Sepharose 4B亲和柱和Sephacry1 S 100分子筛层析纯化了hITF。SDS-PAGE和氨基酸分析证明得到了hITF纯品。以天然hITF为抗原免疫青紫蓝兔制备了抗血清，用纯化的IgG建立了hITF的酶联免疫吸附检测(ELISA)方法，给出了标准工作曲线。应用于纯化过程中样品峰的确立。

关键词: 小肠三叶因子, 酶联免疫吸附检测, 克隆表达

Abstract: An efficient prokaryotic expression hITF system was established in E.coli with pGTF through PCR amplification. The amount of fusion protein GST-hITF was about 15% of the total cellular proteins. The molecular weight of rhITF was 6 kD which was correspondent with that expected. Using hITF as antigen, the antiserum was produced in rabbit. Then enzyme-linked immunosorbent assay (ELISA) was developed for detection and quantitative analysis of human intestinal trefoil factor, and the standard curves were supplied.

Key words: intestinal trefoil factor, enzyme-linked immunosorbent assay (ELISA), clone and expression

中图分类号:

Q516

口如琴,王蔚,李令媛,茹炳根. 人小肠三叶因子的克隆及酶联免疫吸附检测[J]. 北京大学学报（自然科学版）.

KOU Ruqin,WANG Wei,LI Lingyuan,RU Binggen. Clone and Enzyme-Linked Immunosorbent Assay of Human Intestinal Trefoil Factor[J]. Acta Scientiarum Naturalium Universitatis Pekinensis.

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://xbna.pku.edu.cn/CN/

https://xbna.pku.edu.cn/CN/Y2000/V36/I6/777

[1]	杨金水, 高全秀, 李兆胜, 邢冠岚, 袁红莉. 小球藻Chlorella variabilis NC64A二酰甘油酰基转移酶基因的克隆表达与功能研究[J]. 北京大学学报（自然科学版）, 2016, 52(2): 187-192.
[2]	郑军恒；茹刚；茹炳根. 人胎肝金属硫蛋白的分离纯化及酶联免疫吸附检测[J]. 北京大学学报（自然科学版）, 1999, 35(2): 225-229.