人 F3-GRIM19基因的克隆与表达
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孙厚良, 刘 洋, 李甲初,曾昭淳
GRIM19基因;克隆;表达;纯化,人F3-GRIM19基因的克隆与表达,孙厚良，刘洋，李甲初，曾昭淳，,孙厚良,，,CloningandexpressionofhumanF3-GRIM19gene
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孙厚良，刘洋，李甲初，曾昭淳，重庆医科大学生物化学与分子生物学教研室重庆市 400016
孙厚良，男，1975-08-10生，重庆市人，汉族，重庆医科大学生物化学与分子生物学硕士研究生，主要从事肿瘤分子生物学方面的研究.
重庆市教委资助项目，No.1
通讯作者：曾昭淳，400016，重庆医科大学生化与分子生物学教研室.zengzc88@yahoo.com.cn
电话：023-68485398
收稿日期：2005-06-28 接受日期：2005-07-06
Cloning and expression of human F3-GRIM19 gene
Hou-Liang Sun, Yang Liu, Jia-Chu Li, Zhao-Chun Zeng
Hou-Liang Sun, Yang Liu, Jia-Chu Li, Zhao-Chun Zeng, Department of Biochemistry and Molecular Biology, Chongqing University of Medical Science, Chongqing 400016, China
Supported by Program of Educational Administration of Chongqing, No.1
Correspondence to: Dr. Zhao-Chun Zeng, Department of Biochemistry and Molecular Biology, Chongqing University of Medical Science, Chongqing 400016, China. zengzc88@yahoo.com.cn
Received: 2005-06-28 Accepted: 2005-07-06
Abstract
AIM: To clone the cDNA of genes associated with retinoid-IFN-induced mortality (GRIM19), and to construct the F3-GRIM19 expression vector.
METHODS: The total RNA was extracted from normal placenta tissues, and the GRIM19 cDNA was amplified by reverse transcription polymerase chain reaction (RT-PCR). Then the pQE30-F3-GRIM19 expression vector was constructed and sequenced. Thus, the fusion protein of F3-GRIM19 was expressed in E. coli and purified using Ni2+-NTA agarose resin.
RESULTS: After sequencing, the pQE30-F3-GRIM19 contained 580 bp insert, which was consistent with the F3-GRIM19 sequence in GenBank. The expression of the fusion protein (Mr 25 000) was at the highest level 4-5 h after induced by Isopropyl-b-D-thiogalactopyranoside (IPTG), which accounted for 25% in E. coli. After purification, the purity of the target protein was above 90% and the recovery rate was 17.3%.
CONCLUSION: TFL can protect duck liver against inflammation bythe inhibition of DHBV ......
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