摘要
目的:克隆小鼠CD1d1编码区基因。方法:提取小鼠小肠组织总RNA,用RT-PCR技术扩增小鼠CD1d1编码基因,PCR产物连接pGEM-T载体,转化大肠杆菌,用限制性酶切和DNA序列测定对阳性克隆进行鉴定。结果:扩增出一条与预期片段大小相符的特异DNA条带,限制性酶切应显示目的基因片段插入T载体,序列测定表明所获取的基因大小为1011bp。结论:成功克隆了小鼠CD1d1编码基因,为进一步重组蛋白及其抗体的制备奠定了基础。
OBJECTIVE:To clone the gene encoding mouse CD1d1(mCD1d1).METHOD:Total RNA was extracted from the tissue of mouse small intestine.CD1d1 gene was amplified by RT-PCR,and was cloned into pGEM-T vector,which was subsequently transformed 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 agarose electrophoresis.The mCD1d1 gene was inserted into the pGEM-T vector.The result of the DNA sequence showed that a gene fragment of 1011bp was achieved.CONCLUSION:The gene encoding mCD1d1 was cloned successfully,which lays the foundation for further preparation of the recombinant mCD1d1 and its antibody.
出处
《九江学院学报（自然科学版）》
CAS
2008年第3期9-11,共3页
Journal of Jiujiang University：Natural Science Edition
基金
广东省中医药局中医药强省科研基金资助项目(编号1060043)
关键词
CD1D
基因
克隆
鉴定
mCD1d
gene
cloning
identification
