Construction and characterization of the rat neutral amino acid transporter HA － SNAT3 fusion protein

• Main Authors: CHEN Chen, ZHANG Zhou
• Format: Article
• Language: English
• Published: Academic Journals Center of Shanghai Normal University 2014-06-01
• Series: Journal of Shanghai Normal University (Natural Sciences)
• Subjects: neutral amino acid transporter; HA-SNAT3 fusion protein; construction and characterization
• Online Access: http://qktg.shnu.edu.cn/zrb/shsfqkszrb/ch/reader/create_pdf.aspx?file_no=201403001&flag=1&year_id=2014&quarter_id=3

Sodium-coupled neutral amino acid transporter 3 (SNAT3) is a member of SLC38 gene family which primarily expresses in astrocytes in the brain and retina.It transports L-glutamine predominantly and plays an important role in glutamate-glutamine cycle in the brain and liver.In order to easily detect the expression and localization of SNAT3 on the membrane,we constructed a eukaryotic expression plasmid with an HA tag at the N-terminus of rat SNAT3 in the mammalian expression vector p<i>BK-CMVΔ(1098-1300)</i> by PCR and double restriction endonuclease digestion techniques.After transiently transfected the plasmid into human embryonic kidney cells (HEK293T),expression of HA-SNAT3 fusion protein was detected by western boltting.The result suggested that the HA-SNAT3 fusion protein could normally express on the membrane.The plasmid pBK-CMVΔ(1098-1300)-HA-SNAT3 will provide a useful tool for the investigation of structure and function of SNAT3 in the future.