小鼠CD1d2编码区基因的克隆与鉴定

Cloning and identification of the gene encoding mouse CD1d2

目的:克隆小鼠CD1d2编码区基因。方法:提取小鼠胸腺组织总RNA,用RT-PCR技术扩增CD1d2cDNA,PCR产物连接pGEM-T载体,转化大肠杆菌,用限制性酶切反应和DNA测序对阳性克隆进行鉴定,用BLAST软件进行序列分析。结果:扩增出一条特异DNA条带,DNA序列测定表明获取了大小为1008bp的小鼠CD1d2基因编码区基因。结论:成功克隆了小鼠CD1d2编码区基因。

Objective：To clone the gene encoding mouse CD1d2 （mCD1d2）. Methods：Total RNA was extracted from the thymus tissue, mCD1d2 was amplified by RT-PCR and was cloned into pGEM-T vector,which was subsequently tranfomed into E.Coli DH5α, the insertion of gene and the recombinant plasmid were identified by restrict enzyme digestion and DNA sequencing. Results：A specific DNA band was seen by agrose electrophoresis,the result of the DNA sequencing showed that the mCD1d2 coding regeion gene of 1008 bp was obtained. Conclusion：The mCD1d2 coding regeion gene cloned successfully,which laid the foundations for further research on the function of mouse CD1d2.