IL-24融合蛋白表达载体的构建及其对人小肠癌细胞的生长抑制作用
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瞿少刚, 张秀萍, 陈 群, 刘 仿
IL-24;原核表达;GST-IL-24融合蛋白;小肠癌细胞系HIC,瞿少刚,陈群,刘仿,张秀萍,通讯作者:,Constructi
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瞿少刚, 陈群, 刘仿, 广东医学院微生物学及免疫学教研室 广东省湛江市 524023
张秀萍, 广东医学院实验中心 广东省湛江市 524023
瞿少刚, 男, 1971年出生, 湖北省广水市人, 汉族, 2005年武汉大学医学院硕士, 讲师, 主要从事微生物学和免疫学的教学与科研.
通讯作者: 瞿少刚, 524023, 广东省湛江市广东医学院微生物学及免疫学教研室. sgq9528@163.com
电话: 0759-2388583-6 传真: 0759-2284104
收稿日期: 2005-11-09 接受日期: 2005-11-24
Construction of IL-24 gene expression vector and inhibitory effect of IL-24 fusion protein on growth of human intestinal carcinoma cells
Shao-Gang Qu, Xiu-Ping Zhang, Qun Chen, Fang Liu
Shao-Gang Qu, Qun Chen, Fang Liu, Department of Microbiology and Immunology, Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China
Xiu-Ping Zhang, Experiment center, Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China
Correspondence to: Shao-Gang Qu, Master, Department of Microbiology and Immunology, Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China. sgq9528@163.com
Received: 2005-11-09 Accepted: 2005-11-24
Abstract
AIM: To construct a recombinant expression vector of human interleukin-24 (IL-24) gene and express it in E. coli, and to investigate the inhibitory effect of IL-24-GST fusion protein on the growth of human intestinal carcinoma (HIC) cells.
METHODS: The human IL-24 cDNA fragment was amplified from plasmid TRAP-hIL-24 by polymerase chain reaction (PCR), then cloned into the prokaryotic vector pGEX-KG, and expressed inductively as a fusion protein in E. coli. The expressed GST-IL-24 fusion protein was purified via GST-Sepharose 4B Column and identified by SDS-PAGE and Western blot. In vitro cultured human HIC cells were treated with different concentrations of GST-IL-24 fusion protein and GST protein, and the proliferation of HIC cells was measured by MTT assay.
RESULTS: Restriction enzyme digestion analysis showed that the recombinant prokaryotic expression vector pGEX-KG-IL-24 was constructed successfully and expressed stably in E ......
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