人高迁移率族蛋白1基因的克隆和表达

目的：克隆人高迁移率族蛋白1(HMG—1)编码基因，构建重组表达载体并对其诱导表达．方法：通过RT—PCR方法扩增出人外周血单个核细胞中HMG—1的编码基因，克隆于载体pGEM—T easy，测序分析后亚克隆于表达载体pGEx—4T—2，测序证实序列正确后转化大肠杆菌DH5α，经IPTG诱导4h后可表达M，约56000的融合蛋白GST—HMGl．结果：克隆了人HMG—1的编码基因，构建了融合蛋白的重组表达质粒pGEx—HMGl，表达的融合蛋白产物经凝胶薄层扫描检测，占全菌总蛋白的0．23．结论：获得了人HMG—1编码基因及其原核表达产物，对研究人HMG—1的生物学功能及其mAb的制备具有重要意义．

Cloning and expression of the human high mobility group-1 protein code gene

AIM: To clone human HMG 1 code gene, construct the recombinant vector and express its products. METHODS: Human HMG 1 code gene was amplified by RT PCR from human primary peripheral blood mononuclear cells and cloned into vector pGEM T. After sequenced, it was subcloned into prokaryotic expressive vector pGEX 4T 2 .Having induced by IPTG for 4 h, the expression of recombinant M r 56 000 fusion protein GST HMG1 was confirmed by SDS PAGE. RESULTS: Human HMG 1 code gene was cloned. Expressed recombinant plasmid pGEX HMG1 was constructed. The expressed fusion protein was about 0.23 of total bacterial protein by gel thin layer chromatography scanning. CONCLUSION: The human HMG 1 code gene and prokaryotic expression products are obtained. It may be of great significance to the study on the function of human HMG 1 and the preparation of some monoclonal antibody against human HMG 1.