Title: Construction of the mouse T cell immunoglobulin mucin 3( Tim-3) eukaryotic expression vector
Abstract: Objective To construct an eukaryotic expression vector of murine T cell immunoglobulin mucin 3( Tim-3) , and express Tim-3 in the melanoma cell line B16. Methods The murine Tim-3 coding region was amplified by reverse transcription PCR ( RT-PCR) ,using RNA from spleen cells as a template. The Tim-3 cDNA was cloned into the eukaryotic expression vector pTARGET by T-A cloning. The recombinant vector was transfected into the melanoma cell line B16 by Lipofectamine. Tim-3 expression was analyzed by RT-PCR and Western blot. Results The murine Tim-3 expression vector was identified by enzyme digestion and sequencing. RT-PCR and Western blot results showed specific Tim-3 expression in B16 cells transfected with the recombinant vector. Conclusion The Tim-3 coding region was successfully cloned into the eukaryotic expression vector and highly expressed in the melanoma cell line B16.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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