白细胞相关免疫球蛋白样受体-2的克隆、原核表达及纯化

The Cloning, Prokaryotic Expression and Purification of Leukocyte-associated Immunoglobulin-like Receptor-2

以人白细胞相关免疫球蛋白样受体-2(LAIR-2/CD306)cDNA质粒为模板,通过PCR技术获得该蛋白成熟区基因片段,插入表达载体pET28a的多克隆位点,进而将构建的重组质粒导入大肠杆菌BL21(DE3)中进行表达,所得重组蛋白以包涵体形式存在,包涵体经变性、复性及亲合色谱分离后获得重组蛋白。

Human leukocyte-associated immunoglobulin-like receptor-2 （LAIR-2/CD306） was used as template and the mature domain gene was amplified by PCR. The fragment was cloned into the multiple cloning sites of expression vector pET28a. The recombinant plasmid was transformed into E. coli BL21 （DE3） for the expression of LAIR-2. The expressed protein mainly existed in the form of inclusion body. The purified protein was obtained from inclusion body via degeneration, renaturing and affinity chromatography.