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Cloning and expression of tissue inhibitor of metalloproteinases-1 (TIMP-1) gene in Chinese
Shuang-Hu Liu, De-Ming Tan, Jue Hou, Guo-Ling Hu
Shuang-Hu Liu, De-Ming Tan, Jue Hou, Guo-Ling Hu, Department of Infectious Diseases, the Affiliated Xiangya Hospital, Central-South University, Changsha 410008, Hunan Province, China
Supported by: Natural Science Foundation of the Ministry of Health of China, No. 98-1-120.
Correspondence to: Dr. Shuang-Hu Liu, Department of Infectious Diseases, the Affiliated Xiangya Hospital, Central-South University, Changsha 410008, Hunan Province, China. liushuanghu@hotmail.com
Received: November 19, 2002 Revised: November 30, 2002 Accepted: December 2, 2002 Published online: August 15, 2003
AIM
To clone the Chinese gene of TIMP-1 and express the fusion protein MBP-TIMP-1.
METHODS
The fragment of TIMP-1 gene from the liver tissue of indigenous Hunan residents was amplified by RT-nest-PCR and it was cloned, sequenced by gene recombinations techniques. Fusion protein MBP-TIMP-1 was expressed in E.coli, analysed by SDS-PAGE and Western blot and purified by affinity chromatography.
RESULTS
The sequence of cloned Chinese TIMP-1 gene was 624 bp. It is homologous with the reported other human TIMP-1 gene sequence. Fusion protein MBP-TIMP-1 was 66 kd and is of the antigenicity of TIMP-1. The MBP-TIMP-1 was purified by affinity chromatography.
CONCLUSION
The Chinese gene of TIMP-1 was cloned and MBP-TIMP-1 was expressed and purified successfully in vitro. It may be useful to the diagnosis of liver fibrosis.
Key Words: N/A
Citation: Liu SH, Tan DM, Hou J, Hu GL. Cloning and expression of tissue inhibitor of metalloproteinases-1 (TIMP-1) gene in Chinese. Shijie Huaren Xiaohua Zazhi 2003; 11(8): 1164-1167
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