2001•Xinjiang Yike Daxue xuebaoRequires access
The cDNA cloning and sequence about tissue inhibitor of metalloproteinase-1 from human breast cancer
Hao Wen
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Abstract
Objective: In order to expression the cDNA of tissue inhibitor metalloproteionase (TIMP-1) in eukaryotic cell. Method: Total RNA were isolated from human breast cancer cells and cDNA encoding tissue inhibitor metalloproteionase (TIMP-1) was amplified by the reverse transcription PCR(RT-PCR). The cDNA was inserted into vectors PET-19b and the cloning vector PET19bTIMP-1 was constructed. The cDNA was identified by restriction endonucleases and sequenced. Result: It was found to differ in nucleotide as compared to the cDNA sequence(Andeson,et al,1999).This changes cause amino acid substitution at position 361 where a Thr changed into a Ala. This cDNA encodes 207 amino acids whose length is 624bp. Conclusion: The recombinant cloning vector pET19bTIMP-1 was successfully constructed in our Labs. The single gene mutation did not influence the antigen-expression.