中文摘要：

本研究旨在建立一套适合于十字花科黑腐病菌（Xanthomonas campestris pv.campestris 8004,Xcc8004）分泌蛋白质的双向电泳技术（two-dimensional electrophoresis,2-DE）,以便更好的利用蛋白质组学技术鉴定Xcc 8004的分泌蛋白质。我们就MME和XCM2两种培养基对分泌蛋白质的诱导能力、蛋白质提取方法以及样品上样量等方面对2-DE的影响进行了比较,结果表明,XCM2培养基对Xcc 8004分泌蛋白质的诱导能力比MME强。用TCA/丙酮法沉淀蛋白质得到的双向电泳图谱背景清晰,分辨率高。分别采用400μg、300μg、250μg和150μg四个不同的上样量进行双向电泳,结果显示在上样量为250-300μg时（IPG pH 3-10,24 cm）,电泳图谱分辨率最好。综上可知,XCM2培养基比较适合用于Xcc 8004分泌蛋白质的诱导,TCA/丙酮法提取分泌蛋白质为较优方法,250-300μg是较为合适的上样量。

英文摘要：

In order to establish a suitable and efficient two-dimensional electrophoresis（2-DE） technique for identifying the secreted proteins of Xanthomonas campestris pv. campestris 8004, the extracellular proteins on inducing media, MME and XCM2, different protein extraction methods, and loading amount on 2-DE profile were compared in this research. The results showed that XCM2 is more suitable for inducing extracellular proteins than that of MME, and clearer 2-DE maps can be observed while the proteins were precipitated by the TCA/acetone treatment. Furthermore, different loading amount（400 μg, 300 μg, 250 μg and 150 μg） of the proteins extracted by the method of TCA/acetone for 2-DE were used to optimize the conditions of sample preparation. As a result, the resolution and the quality of the 2-DE maps were most improved for proteins loading amount of 250μg to 300 μg separated in 24 cm IPG strips（pH 3-10）. To sum up, XCM2 is suitable for inducing extracellular protein,TCA/acetone precipitation is much more suitable for protein extraction and 250-300 μg is the optimum loading amount of 2-DE for secreted proteins of Xanthomonas campestris pv. campestris.