双向电泳联用质谱技术研究棉苗对细极链格孢菌蛋白激发子诱导的应答

Analysis of the Inducing Responsiveness in Cotton Against Protein Elicitor from Alternaria tenuissima Using Two-dimensional Electrophoresis and Mass Spectrometry

采用双向电泳和质谱技术分析了细极链格孢菌蛋白激发子对棉苗诱导的全蛋白质组,建立二者间的差异表达图谱,并对差异蛋白进行了鉴定及分析归类。结果表明,蛋白质分子质量在10～100kD之间、等电点3～10范围内,每块胶分离到约600个蛋白质点。得到了大约53个差异蛋白点,其中有31个为上调蛋白,22个为下调蛋白。对PEAT诱导后表达量明显增加的7个上调蛋白点用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOFMS)进行肽质量指纹谱的分析,并通过检索不同的数据库进行蛋白质鉴定与功能预测,获得了4个蛋白的肽质量指纹图谱,经数据库检索鉴定分析,GH-1、GH-3、GH-6推测为核酮糖1,5-二磷酸羧化酶基因家簇,GH-7推测为S-腺苷一蛋氨酸合成酶。

Proteins extracted from the cotton seedling induced by protein elicitor from Alternaria te nuissima were separated by two dimensional polyacrylamide gel electrophoresis （2-DE） and Mass Spectrometry. Reference maps were obtained with a good reproducibility. After analyzing with 2-DE software, approximately 600 proteins per 2-DE gel were isolated with molecular weights ranging from 10 kD to 100 kD and PI ranging from 3 to 10. About 53 spots expressed differently, with 31 at a higher level and 22 at a lower level. Seven stronger expressed proteins of high levels observed in a seconda ry PEAT gel were excised and identified using peptide mass fin-reprinting and matrix assisted laser desorption ionization time-0f-flight mass spectrometry. Four identified proteins and their putative functions were presented. Three of them, spot no. 1,3,6 were identified as ribulose-1,5 bisphosphate carboxylase,and spot no. Seven was identified as S-adenosyl L methioine sythetase.