• Original article (Basic research) • Previous Articles     Next Articles

Expression, purification and characterization of human inhibitory receptor TIGIT in E.coli

CHEN Xiao-yun1, ZHOU Ai-wu2, YE Wei1   

  1. 1. Department of Preventive and Pediatric Dentistry, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; 2. Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

  • Online:2017-02-28 Published:2017-02-28
  • Supported by:

    Foundation of Shanghai Ninth Peoples’s Hospital, Shanghai Jiao Tong University School of Medicine, 2012-04


Objective · To prepare recombinant human TIGIT protein in E.coli and characterize its ability in binding Fusobacterium nucleatum (Fn). Methods · The gene of immunomodulatory protein of human TIGIT was amplified and cloned into pGEX4T2, and recombinant plasmid was transformed into E.coli BL21 (DE3) for GST-TIGIT fusion proteins were purified by the GST affinity chromatography and the interaction between GST-TIGIT fusion protein and Fn was tested by a pulldown assay. Results · Recombinant GST-TIGIT fusion protein expressed successfully in E.coli and was purified to homogeneity by GST affinity column. This protein could specifically bind to Fn, but not Lactobacillus acidophilus. Conclusion · High purify and activity of human GST-TIGIT fusion protein can be achieved by the prokaryotic expression system, and the adhesion between this protein and Fn has been preliminarily explored, which provides basis for further characterize interaction between them.

Key words: TIGIT, clone, expression, purification, Fusobacterium nucleatum, adhesion