人脂联素在大肠杆菌中的可溶性表达

为了提高重组人脂联素在大肠杆菌中的可溶性,构建和表达了增溶标签与人脂联素的融合表达栽体.这些标签包括大肠杆茵硫氧还蛋白和NusA蛋白、酿酒酵母SUMO蛋白和噬热海洋茵Thermotoga maritime 的红素氧还蛋白.结果显示,所有的人脂联素融合蛋白在大肠杆茵中都获得了高水平的表达,但可溶性存在很大差异,说明不同融合标签对人脂联素可溶性表达的促进作用不同.其中,红素氧还蛋白与SUMO蛋白组合一起对人脂联素可溶性的增强作用最为显著,其相应的融合蛋白几乎全部可溶.另外,通过酵母SUMO/Ulp1反应,经His标签亲和层析纯化得到的人脂联素融合蛋白能被有效酶切,加工为人脂联素成熟肽产物.

Soluble expression of human adiponectin in Escherichia coli

In order to improve the solubility of recombinant human adiponectin in Escherichia coli, con structs for expressing human adiponectin protein fusions with a few of solubility- enhancing tags were generated. These tags include two E. coli proteins, Thioredoxin and NusA, SUMO protein of yeast Saccharomyces cerevisiae and Rubredoxin from thermophilic oceanic bacterium, Thermotoga maritime.The results demonstrate that all human adiponectin fusion proteins were highly expressed, but with remarkably different solubility, implying that these fusion tags differentially improve the soluble expression of human adiponectin. Therein, the combinational tag of Rubredoxin and SUMO exhibited the highest solubility-enhancement, and its correlated fusion protein was almost completely soluble. Furthermore, by using yeast SUMO/Ulpl reaction, human adiponectin fusion proteins purified by His-tag affinity chromatography were efficiently processed into the mature adiponectin by enzymatic cleavages.