人cd 226 分子胞膜外区d1 和d2 真核表达载体.pdf

23 5 2007 9 IMMUNOLOGICAL JOURNAL Vol. 23 No. 5 Sep. 2007 499 ( ) [ ]100028861 2007 0520499204 CD226 D1 D2 3 ( ,,, ,,,, , 710032) ( ) ( ) [ ] CD226 1 D1 2 D2 , CD226 D1 D2 ,pSecTag2B2Fc ,DNA , 293T ,, SDS2PAGE Western blot PCR CD226 D1 D2 ,pSecTag2B2Fc , 293T ,Western blot 293T hCD226D12Fc hCD226D22Fc ,, hCD226D12Fc hCD226D22Fc ,, [ ] CD226 ;; [ ] R392. 11 [ ] A Construction and expression of human CD226 extracellular domain 1 and domain 2 gene eukaryotic expression vectors and identification of the fusion proteins DEHG Hu2ping , ZHUANG Ran , J IA Wei , ZHANG Yun , ZHANG Yuan , HU Yi , ZHANG Xin2hai , J IN Bo2quan ( Department of Immunology , Fourth Military Medical University , Xi an 710032 , China) [ Abstract ] Objective To construct and express eukaryotic expression vectors of domain 1 and domain 2 in the extracellular region of human CD226 and identify their expressed fusion proteins. Methods The hCD226D1 and hCD226D2 genes were amplified from the full length CD2262flag and cloned to pMD182T vector by PCR. The recombinant plasmids were digested and inserted into the same site of expression vector pSecTag2B to construct pSecTag2BΠhCD226D1 and pSecTag2BΠhCD226D2. The recombinant plasmids were digested and subjected to gene se2 quencing before recombinant expression vectors were transfected into 293T cells. The expressed fusion proteins hCD226D12Fc and hCD226D22 Fc were purified by WT6 conjugate