人粘膜血管定居因子／GST融合基因表达载体的构建与表达

目的 构建hMAdCAM-1/GST融合基因表达载体并进行表达。方法 采用PCR技术扩增hMAdCAM-1cDNA5‘端615bp的基因片段，将其插入pGEM-T质国。经全自动序列分析仪测序证实后，再亚克隆至表达载体pGEX-2T，转化大肠杆攻DH5a。结果 SDS－PAGE检测显示，表达出相对分子量52000u的融合蛋白，占菌体总蛋白的31％左右。Westerm blot分析表明，表达蛋白能与抗hMAdCAM-1多抗特异性结合。结论 hMAdCAM-1/GST融合基因表达载体的构建和表达，为深入研究MAdCAM-1提供了材料。

Construction and expression of recombinant hMAdCAM-1/GST fusion gene expression vector

ObjectiveTo construct and express a recombinant expression vector bearing hMAdCAM-1/GST fusion gene. MethodshMAdCAM-1 cDNA 5'-end 615 bp fragment encoding hMAdCAM-1 Ig-like domains was amplified by PCR from plasmid pUC21/hMAdCAM-1 and then inserted into pGEM-T plasmid. Determined by auto-sequencing, the target gene fragment was cloned into prokaryotic expression vector pGEX-2T in fusion form and transformed into E.coli DH5a. ResultsSDS-PAGE analysis showed that a new protein band with molecule weight of 52 000 u appeared as the expected size, representing 31% of the total bacterial protein. Western-blot indicate that the expressed fusion protein could bind with antiserum against hMAdCAM-1 specifically. ConclusionThe successful construction and expression of recombinant vector pGEX-2T-MAd might provide material for the further research about hMAdCAM-1.