水稻OmR40c1蛋白的表达纯化及其与血红细胞的凝集作用

Fusion Expression and Purification of a Rice Protein Om R40c1 and its Agglutination Effect

目的:在大肠杆菌中融合表达水稻蛋白Om R40c1,并探讨其与红细胞的凝集效果,为进一步研究Om R40c1在体外的抑菌实验做准备。方法:PCR扩增目的基因,经Bam HⅠ及NotⅠ双酶切,连入表达载体p GEX-6p-1构建成重组质粒,测序正确后转入大肠杆菌BL21,优化诱导表达条件;纯化重组Om R40c1蛋白,并分析其与红细胞的凝集效果。结果:获得原核表达的Om R40c1蛋白,重组Om R40c1在低温(4℃)条件下可与兔血红细胞发生凝集作用。结论:原核表达体系适合水稻Om R40c1蛋白的融合表达。

Objective： To express and purify fusion rice protein OmR40c1 and explore its agglutination effect.Methods： OmR40c1 gene was amplified by PCR from Oryzae meyeriana c DNA, a wild rice in south China, andwas cloned into p MD19-T Blunt Simple vector. The purified plasmid was digested with Bam HⅠ/NotⅠ and theOmR40c1 gene was cloned into the corresponding sites of p GEX- 6p- 1. Recombinant plasmid p GEX- 6p- 1-OmR40c1 was transformed into E.coli BL21 to express OmR40c1-GST fusion protein, with optimize two factors.Then, the agglutination effect analysis was carried out between OmR40c1 and rabbit red cells. Results： Fusion pro-tein OmR40c1 was successfully expressed in E.coli BL21 and purified by using affinity chromatographic separation.Mixture of OmR40c1 and red cells can be aggregated under 4℃. Conclusion： OmR40c1 is a lectin and can beexpressed in Escherichia coli BL21.