Expression and identification of rats glutamine transporters EGFP-SNAT3 fusion protein

Journal of Shanghai Normal University (Natural Sciences)

ZHANG Zhou¹  MIN Weiyong²  LI Min² 

[1] College of Life Science and Biopharmacy,Shenyang Pharmaceutical University;Collegeof Life and Environmental Sciences,Shanghai Normal University;

关键词: enhanced green fluorescent protein;    EGFP-SNAT3 fusion protein;    plasmid construction;    expression and identification;   

DOI  :  10.3969/J.ISSN.100-5137.2014.02.011

来源: DOAJ

【 摘 要 】

The sodium-coupled neutral amino acid transporter 3(SNAT3) encoded by the SLC38 gene family is a member of the System N.It plays an important role in Glutamate-Glutamine shuttle in the brain and glutamine transportation in the membrane of liver cells.In order to detect expression and localization of SNAT3 on the membrane conveniently,in this study,an enhanced green fluorescent protein(EGFP) sequence was constructed to the N terminus of SNAT3 by PCR amplification,enzyme digestion and ligation.After the recombinant eukaryotic expression plasmid of pBK-CMV(Δ［1098-1300］)-EGFP-SNAT3 was transiently transfected into HEK293T cells mediated by liposomal for 36 hours,expression and localization of EGFP-SNAT3 fusion protein were detected by laser scanning confocal microscope and Western blot.The results showed that EGFP-SNAT3 expressed and localized correctly on the membrane.The recombinant plasmid of EGFP-SNAT3 constructed successfully may contribute to the further study of structure and function of SNAT3 in the future.

【 授权许可】

Unknown