摘要
目的 克隆人CD13 7分子的编码序列 ,将其重组入真核表达载体 ,并表达于COS 7细胞。方法 从活化的人T细胞cDNA文库中以PCR方法克隆编码人CD13 7的编码序列 ,对其序列进行测定。将测序正确的CD13 7编码序列克隆入真核表达载体PCI neo ,构建重组表达载体。采用脂质体法转染COS 7细胞 ,用流式细胞仪检测CD13 7分子的表达。结果 PCR方法扩增出一 80 0bp左右的基因片段 ,插入PCI neo构建重组表达载体后 ,经序列测定证实扩增的片段为人CD13 7编码序列。将重组子转染COS 7细胞 ,流式细胞仪检测显示有 2 7.11%的细胞表达人CD13 7分子。结论 成功地克隆并在COS 7细胞中表达了CD13 7分子。
Objective To clone full length human CD137 encoding sequence and to express functional CD137 molecule on the COS 7 cells. Methods PCR technique was used to clone the full length DNA encoding CD137 molecule from human T cell cDNA library. The PCR product was digested and inserted into PCI neo vector and sequenced. The right recombinant was transfected into COS 7 cell with lipofectamine reagent. The expression of CD137 molecule on the membrance of COS 7 cells was detected with FACS. Results A fragment about 800bp was cloned from human T cell cDNA library, and was subsequently digested and cloned into PCI neo vector. Sequence analysis demonstrated that the fragment cloned fits in with the published CD137 encoding sequence. FACS revealled that 27.11% cells expressed human CD137 molecules. Conclusion CD137 is cloned successfully and expresses in COS 7 cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2002年第11期1343-1345,共3页
Journal of Third Military Medical University
