细胞黏附分子1(ICAM-1)ScFv基因原核载体的构建及表达

Construction of prokaryotic expression vector and expression of ICAM-1 ScFv gene

应用PCR技术扩增ICAM-1ScFv基因,经纯化测序,得到约750bp的核苷酸片段,并构建了原核重组表达质粒pET20b-ICAM-1ScFv。将重组质粒转化至表达菌BL21(DE3)中,诱导后的SDS-PAGE分析显示,pET20b-ICAM-1ScFv表达量占菌体蛋白总量的18.4%。本试验将重组蛋白的表达条件进行优化,结果表明,ICAM-1ScFv蛋白在pET20b表达载体中的最佳表达条件为37℃诱导5h。

The（ICAM-1）ScFv gene of 750 bp were amplified by PCR.The recombinant expressing plasmid pET20a-ICAM-1 ScFv were obtained and recombinant proteins was expressed in BL21（DE3）.Analysis of SDS-PAGE and thin layer scanning results showed that amount of expressed recombinant proteins was 18.4% in total lysis protein of bacteria.The expression condition was improved and the result indicates that the proper factor was 37℃,induced 5 h with IPTG.