氮胁迫条件下稻瘟病菌菌丝体蛋白质组学研究

Preliminary study of mycelium protein in Magnaporthe oryzae using two-dimensional electrophoresis under nitrogen starvation

采用改进的三氯乙酸/丙酮法,对稻瘟病菌菌丝体蛋白质进行双向电泳,获得了高分辨率电泳图谱,较改进前三氯乙酸/丙酮法蛋白质点数量平均提高687个点。在等电点pI 3～10和4～7范围内,电泳图谱上可分别检测到(500±40)及(984±70)个蛋白点。利用该方法对氮胁迫条件下稻瘟病菌菌丝体蛋白质进行双向电泳,在pI 4～7范围内电泳图谱上检测到(1 169±90)个蛋白点。通过加强对样品的清洗结合丙酮对蛋白质在水化液中的二次沉淀,能较好地除去样品中杂质而保证电泳质量。优化后的方法可用于稻瘟病菌菌丝体蛋白质组的研究,并获得了更加理想的效果。

The high-resolution two-dimensional electrophoresis （2-DE） maps of mycelium protein in Pyricularia oryzae were obtained through improved TCA/acetone method and the spots increase 687. Spots of （500±40） and （984± 70） were detected on the 2-DE maps with pI 3--10 and pI 4--7 respectively. Spots （1 169±90） were also detected on the 2-DE maps with pI 4--7 of mycelium protein in P. oryzae after nitrogen starvation. Impurities of protein samples could be removed effectively after enhanced sample cleaning and the secondary protein precipitation from rehydration solution using acetone. This study provides a convenient and efficient technique for mycelium protein detection in P. oryzae analysis by using 2-DE.