昆明小鼠Tim-2基因的单个核苷酸变异的研究

Study on the gene variantion of T cells immunoglobulin domain and mucin domain protein-2 in km mouse

目的:研究T细胞免疫球蛋白域黏蛋白域蛋白(T cell immunoglobulin-and mucin-domain-containing molecules,Tim)基因家族的成员之一--Tim-2基因在昆明鼠体内的单核苷酸多态性,以提示昆明鼠在肿瘤学和免疫学的相关试验中有可能出现的与国际通用的其他品系小鼠(如BALB/c小鼠)不同的生物学特性。方法:取不同来源的5只昆明小鼠的肝组织,提取总RNA后逆转录得到cDNA,使用特异性引物通过PCR扩增得到Ti m-2基因,克隆后测序,然后比较测序图谱。结果:测序结果与Genebank中的Ti m-2序列相比,451(TCC→GCC:丝氨酸→丙氨酸),502,504(CCA→TCT:脯氨酸→丝氨酸),586,587(GAT→TCT:天冬氨酸→丝氨酸)。其他变异位点无氨基酸改变。结论:提示昆明小鼠与国际通用的其他品系小鼠(如BALB/c小鼠)Ti m-2基因所编码的蛋白质序列有差异,在使用昆明小鼠进行免疫学相关的试验研究的过程中应该注意到由于这种不同所造成的对实验结果的影响。

Objective: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. Method: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.