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Expression and Identification of Rats Glutamine Transporters SNAT2-EGFP Fusion Protein
Meng Wen, Wang Han, Dong Xiaoyun, Li Yang, Zhang Zhou*
College of Life and Environmental Science, Shanghai Normal University, Shanghai 200234, China
Abstract: Sodium-coupled neutral amino acid transporters (SNATs) in the central neural system played an important role in transporting small， neutral amino acids， such as glutamine and alanine， across cellular membranes. In order to detect expression and localization of SNAT2 on cell membranes conveniently， an EGFP protein sequence was subcloned into SNAT2’s C-Terminus in the eukaryotic expression vector pBK-CMV-(Δ[1 098-1 300])-SNAT2-myc. After pBK-CMV-(Δ[1 098-1 300])-SNAT2-EGFP was transiently transfected into HEK293T cells， expression of SNAT2-EGFP was detected by laser scanning confocal microscope and Western blot using anti-GFP antibody. The results showed that SNAT2-EGFP was successfully expressed and localized on cell membranes. The eukaryotic expression plasmid pBK-CMV-(Δ[1 098-1 300])-SNAT2-EGFP constructed successfully is an effective tool for studying structure and function of SNAT2 in the future.
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