甘蔗叶片蛋白质组双向电泳技术优化

Optimization of Two-dimensional Polyaneylamide Gel Electrophoresis for Proteome of Sugarcane Leaves

为了建立适合甘蔗叶片蛋白质组研究的双向电泳技术,对甘蔗叶蛋白质的溶解方法、IEF电泳、上样量等关键步骤进行了优化.结果表明:裂解液中有2 mmol/L的硫脲才能更充分地溶解蛋白;上样量1.0 mg时得到质量较好的凝胶图谱;甘蔗叶片蛋白质主要分布在pH4～7范围.通过对甘蔗叶片蛋白双向电泳技术的优化,提高了双向电泳图谱分辨率和蛋白点数,建立了一套适用于甘蔗叶片蛋白质组分析的双向电泳(2-DE)方法,为甘蔗糖代谢相关的差异蛋白质组学研究提供参考.

A two-dimensional polyareylamide gel electrophoresis（2D-PAGE） system was established for proteomic study of sugarcane（Saccharum officinarum L.） leaves.The results show that the system containing lysis buffer with 2 mmol/L thiourea for complete dissolution of proteins,loaded proteins 1.0 mg for good protein patterns,sugarcane leaf mainly distributed in the pH 4～7 range.As the optimization of the experimental conditions on two-dimensional electrophoresis,the useful information can be provided for the proteomics research of sugancane leaf during the period of glucose metabolism accumulation.