汾酒大曲宏蛋白质组的制备与双向电泳技术的建立

采用TCA-丙酮沉淀法、丙酮沉淀、硫酸铵等沉淀法制备汾酒大曲的宏蛋白组样品,并用双向电泳来检测制备效果,结果表明：TCA-丙酮沉淀法制备的样品经电泳分离后,减轻了杂质干扰,其2-DE图谱中竖条纹干扰较少,且获得的蛋白点形状规则、清晰且无明显重叠现象,优于其它两种方法;经过两次水化液溶解的蛋白样品在等电聚焦时能保持4000 V较高电压;上样量为300μg左右的粗蛋白溶于二次水化液能得到点数更多、分辨率高的电泳图谱。建立了汾酒大曲宏蛋白质组的双向电泳体系,为汾酒品质研究奠定基础。

The preparation of metaproteome of Fen jiu daqu and establishment of two-dimensional electrophoresis technology

Metaproteomics samples of Fen jiu daqu were prepared by using TCA-acetone precipitation,acetone precipitation and ammonium sulfate precipitation,then analyzed with two-dimensional electrophoresis. The result showed TCA acetone precipitation was superior to that prepared by the other two methods. In 2-DE map,the interference of vertical stripes decreased,the proteins dots were clear and had regular shape without significant overlap. After dissolved samples in hydration solution twice,the voltage during isoelectric focusing could remain 4000 volts. When the sample loading was about 300ug,the 2DE map displayed more spots and the resolution was higher. This study established the 2-DE methodology for metaproteomics from Fen jiu Daqu.