Khabibullina N F, Liukmanova E N, Kopeina G S, Shenkarev Z O, Arsen'ev A S, Dolgikh D A, Kirpichnikov M P
Bioorg Khim. 2010 Sep-Oct;36(5):654-60. doi: 10.1134/s1068162010050080.
The cell-free expression system based on bacterial extract S30 from E. coli for production of the transmembrane domain of human receptor tyrosine kinase ErbB3 (residues 632-675) was developed. The synthesis of the domain in the soluble form in the presence of detergents and in the form of the translation mixture precipitate was studied. The protocols of purification of the recombinant domain obtained by both methods were developed. The final yield of target protein in optimal conditions was 1.8-2.0 mg per 1 ml of translation mixture.
开发了基于大肠杆菌细菌提取物S30的无细胞表达系统,用于生产人受体酪氨酸激酶ErbB3的跨膜结构域(第632 - 675位氨基酸残基)。研究了该结构域在去污剂存在下以可溶形式以及以翻译混合物沉淀形式的合成情况。制定了通过这两种方法获得的重组结构域的纯化方案。在最佳条件下,每1毫升翻译混合物中目标蛋白的最终产量为1.8 - 2.0毫克。