红色红曲菌蛋白质组双向凝胶电泳体系的建立

Establishment of Two-dimensional Electrophoresis System for Proteome of Monascus ruber

红曲菌是一种具有较高食用和药用价值的丝状真菌,能够产生红曲色素、莫纳可林K等多种生理活性物质。通过分析比较培养基、蛋白质裂解液组成以及水化上样条件对双向电泳结果的影响,建立了红色红曲菌蛋白质组的双向凝胶电泳体系,为从蛋白质水平研究红曲菌及其次级代谢产物的生物合成提供依据。结果表明:用YES培养基培养红色红曲菌6 d,TCA-丙酮法提取菌体总蛋白质,蛋白质裂解液组分为8mol/L尿素,2mol/L硫脲,4%CHAPS,1%DTT和2%Bio-lyte,可获得蛋白质样点数量多,清晰度高的双向电泳图像,为进一步研究红曲菌蛋白质组奠定了基础。

Two-dimensional electrophoresis （2-DE） is an important technique in proteomics research. The 2-DE system for proteome of Monascus tuber was established by comparing and analyzing the infection caused by different kinds of mediums, lysis buffer and the condition of rehydration. By cultivating the Monascus ruber with YES for 6 days, extracting total protein by TCA-acetone, lysis buffer with 8 moL/L urea, 2 mol/L thiourea, 4 % CHAPS, 1% DTT and 2 % Bio-lyte, an ideal 2-DE map with higher resolution and better legibility was obtained, which laid a foundation for the further studies on proteome of Monascus tuber.