人白介素6受体功能区片段在E．coli中的表达

通过ＤＮＡ体外重组技术，以ｐＥＴ－３ｂ为表达载体，构建了重组表达质粒ｐＥＴ－６Ｒ（Ｂ）和ＰＥＴ－６Ｒ（Ｂ）４，分别编码２８ｋＤ的ｈＩＬ－６Ｒ配基结合区片段及其５３ｋＤ的二联体蛋白，并为酶切分析和ＤＮＡ序列分析所证实。ＳＤＳ－ＰＡＧＥ分析表明，含有重组表达质粒的菌株可分别表达出２８ｋＤ的蛋白ｒＩＬ６Ｒ－２８和５３ｋＤ的ｒＩＬ６Ｒ－５３。重组蛋白分别占菌体总蛋白的４５％和２９％左右。重组蛋白主要以包涵体形式存在，Ｗｅｓｔｅｒｎ印迹表明重组蛋白具有ＩＬ－６Ｒ的抗原性。

Expression of the Functional Fragment of Human Interleukin 6 Receptor in E. coli

Two recombinant expressing plasmids pET-6R (B) and pET-6R (B)4, encoding the ligand-binding region (28 kD) of hIL-6R and its dimer (53 kD), respectively, were constructed. They were confirmed by endonuclease digestion and DNA sequencing. SDS-PAGE analysis suggested that bacteria containing recombinant plasmids could produce two proteins,rIL6R-28 of 28 kD and rIL6R-53 of 53 kD, respectively. The recombinant proteins amounted to 45% and 29% of total bacteria proteins. The recombinant proteins were mainly recovered as inclusion bodies. Western blotting indicated that recombinant proteins could react with antibodies against hIL-6R.