极低密度脂蛋白受体结合功能片段的克隆与表达

CLONING AND EXPRESSION OF VERY LOW DENSITY LIPOPROTEIN RECEPTOR FRAGMENTS WITH BINDING FUNCTION

建立具有正常极低密度脂蛋白(VLDL)受体结合功能的小分子受体片段模型并借助此模型研究VLDL受体与配体结合的作用机制。PCR扩增获得LBR1-3和LBR1-8两个DNA片段并将其克隆至原核表达载体pET-28a(+),转化大肠杆菌菌株Rosetta(DE3),利用IPTG诱导蛋白表达。表达产物经N i+-NTA亲和层析柱纯化,为VLDL受体的进一步功能与应用研究,特别是针对该受体的配体识别、结合作用机制研究奠定了基础。

The purpose of the paper is to build a module of small molecular receptor fragments eharaetering normal VLDL - R binding function and study the mechanism of VLDL - R binding to ligand. The genes of LBR1- 3 and LBR1-8 were amplified by PCR and the products were inserted into plasmid pET28a （ ＋ ） seperately. The recombinant plasmids were transformed into competent E. coli strain： Rosetta. Recombinant proteins were induced by IPTG. The fusion proteins were purified of Ni^＋ - NTA chromatography. These results lay the foundation of the receptor＇ s structure and apphcation study, especially for its hgand recognition and binding characteristic.