人粒细胞集落刺激因子受体膜外区在昆虫细胞中的表达

以胎盘组织提取的mRNA为模板，RTPCR扩增人粒细胞集落刺激因子(G-CSF)受体膜外区CRH的cDNA片段，克隆于供体质粒pF_ASTB_AC1，与杆状病毒表达载体Bacmid同源重组后，转染昆虫细胞SF9，获得重组杆状病毒并证明了CRH的高效表达。表达产物经G-CSF亲和层析进一步纯化，纯度可达90%以上。受体竞争性结合实验结果表明，该表达产物能特异性结合G-CSF，具有较高的亲和力(Kd=3.8nmol/L)。

Expression of the CRH Region of G CSF Receptor in Insect Cell

The CRH cDNA of G CSF receptor gene was cloned into the plasmid pFASTBAC. After transformed the E.coli DH10BAC, recombinant baculovirus genome which contained CRH cDNA was obtained. By transfecting the insect cell SF 9 with the recombinant baculavirus genome, the recombinant baculavirus was obtained. SDS PAGE and Western blot analysis of the infected cell lysit showed that CRH was expressed in the SF 9 cell. The result of the receptor binding experiment also proved that the expression product purfied by the G CSF affinity column can specifically bind the 125 IG CSF with the Kd of 3.8nmol/L.