烟草烟碱转化相关蛋白质筛选分析研究

为了研究烟草中烟碱转化机理，应用比较蛋白组学方法研究了高烟碱烟草中烟碱转化相关蛋白质表达情况.采用双向电泳联用质谱技术比较了实验组高烟碱转化烟草叶片和对照组野生型烟草叶片的蛋白质差异，获得了分辨率和重复性较好的双向电泳图谱.选取了34个差异蛋白质点，采用MALDI TOF TOF MS进行肽质谱指纹图分析，最终有12个蛋白质点得到了可靠鉴定，其中在实验组中相对下调的蛋白有7个，相对上调的蛋白有5个.通过比对蛋白质组库数据，发现这些差异表达的蛋白质主要是参与碳水化合物代谢、能量代谢等功能，而且亚细胞定位主要是在叶绿体和线粒体,表明这些蛋白表达水平与烟碱转化具有密切关系，为研究高烟碱转化率烟叶的形成机理提供了新的依据.
To study the mechanism of tobacco nicotine conversion,the expression of nicotine conversion related proteins of high tobacco nicotine conversion was explored by differential proteomics. Two dimensional electrophoresis combining MS spectrometry were used to compare the expression of the proteins between high nicotine conversion tobacco (experimental group) and wild type tobacco (control group). After obtaining the distinguishable and repeatable 2 D pictures,there were 34 different proteins spots between the two tobacco plant leaves.12 different protein spots were then identified by the Matrix Assisted Laser Desorption/ Ionization time of Flight Mass Spectrometry Mass Spectrometry (MALDI TOF TOF MS),including 7 protein spots down regulated in white flower leaves (experimental group),and 5 protein spots up regulated. In addition to comparison analyses with proteomic database,these proteins are involved in carbohydrate,energy metabolism and so on. Most of them are chloroplasts or mitochondria proteins. In a whole,these differential expression proteins were closely related with tobacco nicotine conversion,which can provide another pathway for finding the mechanism of tobacco nicotine conversion.