Title: Mutagenesis of human monocyte-chemoattractant protein-1 by recombinant PCR
Abstract: Objective: To construct 7ND CDNA-the deletion mutant of human monocyte chemoattractant protein-1 (MCP-1) cDNA by recombinant PCR. Methods: Using pBluescript-hμ MCP-1 as templates, two sequences before and after the mututant site were amplified with two pairs of synthetic primers. Recombinant PCR introduced the deletion mutant by linking the two sequences, and the production 7ND cDNA was cloned into T vector for further cloning. Then 7ND cDNA was inserted into plasmid pcDNA3.1 to construct 7ND eukarytic expressing vector. Results: A recombinant plasmid pcDNA3.1-7ND expressing human MCP-1 cDNA mutant was successfully constructed. The analysis of sequence proved that 7ND has a length of 342 bps and the N-terminal amino acids of 2 through 8 of human MCP-1 lacked. Conclusion: The result indicate that recombinant PCR is a very effect and reliable method for gene mutant.
Publication Year: 2004
Publication Date: 2004-01-01
Language: en
Type: article
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