Construction of Eukaryotic Expressing Vector of Human TLT-2 and Establishment of Stable Transfectant L929 Cell Line

Objective To construct eukaryotic expressing vector of human trem-like transcript 2(TLT-2) gene and transfect L929 cells so as to establish stable L929 cell line.Methods cDNA fragment encoding TLT-2 was obtained from human lymphocyte cells by reverse transcription.The TLT-2 gene was then designed with the restiction endonucleases XhoI and EcoRI and inserted into corresponding region of pIRES2-EGFP vector.And then the recombinant pIRES2-EGFP/TLT-2 was transfected into L929 cells by lipofectanime TM2000.After screening culture by G418,stable transfected L929 cell line was established,and the expression of TLT-2 was identified by FCM,RT-PCR and Western blot.Results The eukaryotic expressing vector pIRES2-EGFP/TLT-2 was constructed,stable transfected L929 cell line was established,and TLT-2 gene was expressed successfully.Conclusion The construction of eukaryotic expressing vector pIRES2-EGFP/TLT-2 and the development of stable transfected L929 cells have provided a solid experiment foundation for further studies of TLT-2 fuction.