Expression And Localization Of BALB/c Mouse T Cell Immunoglobulin And Mucin Domain Containing Molecules-2 In L929 Cell Line And Facilitate The Ferritin Endocytosis

PartⅠExpression and localization of BALB/c mouse T cell immunoglobulin and mucin domain containing molecules-2 in L929 cell line and facilitate the ferritin endocytosisObjective:To get BALB/c mouse T cell immunoglobulin and mucin domain containing molecules-2 encoding region, and construct eukaryotic expression vector, express efficiently protein in mouse cell line, and study the functions.Methods:Tim-2 gene of BALB/c mouse was obtained by RT-PCR method, and then the gene segments were inserted into pEGFP-N1 vector after T-A clone and assessment. L929 cells were transftected by DOSPER Liposomales Transfektionsreagenz. The expression of Tim-2 gene was demonstrated by fluorescence microscope. Finally, stimulated the transfected cells by ferritin.Result:The Tim-2 gene was efficiently expressed in L929 cells. The fluorescence microscope results indicated the expressed protein localized in membrane system and facilitated the ferrtin endocytosis.Conclusion:Tim-2 gene of BALB/c mouse could express efficiently in mouse cell line and facilitated the ferrtin endocytosis. PartⅡStudy on the gene variantion of T cells immunoglobulin domain and mucin domain protein-2 in km mouseObjective:To investigate the single nucleotide polymorphism (SNP) sites of T cells immunoglobulin domain and mucin domain protein-2 (Tim-2) in km mouse and hint the different bionomics between the km mouse and BALB/c mouse in the oncology and immunologic experiments.Methods:Firstly, obtain the hepatic tissues and extract the total RNA of five km mouse respectively which from different experimental animal centers. Secondly ,Use specific primers to amplificate the objective gene by reverse transcription and polymerase chain reaction (RT-PCR). Finally , sequencing after T-A cloning and compare the atlas.Result:There have some gene variantions in Tim-2, such as: 451(TCC→GCC: Ser→Ala),502,504(CCA→TCT: Pro→Ser),586,587(GAT→TCT: Asp→Ser)。The other variation sites have no amino acids changes。Conclusion:There are some variations of Tim-2 protein between the km mouse and BALB/c mouse. The studies should observe the effect that caused by the variations in the oncology and immunologic experiments.