黑暗条件下木薯叶绿体比较蛋白质组学初步研究

摘要

The chloroplast of the cassava cultivar ‘South China 8’ ( SC8) was used to extract the chloro⁃plast protein with an improved phenol(BPP) method,and these proteins expressed profiles under dark condi⁃tion were determined with 1⁃DE and 2⁃DE.The obtained typical 2⁃DE gels were analyzed by ImageMaster soft⁃ware to determine the differential expressed protein spots,and some of them were manually excised for protein in⁃gel digestion and subsequently identified by MALDI⁃TOF MS. The main results included:high quality pro⁃teins were obtained from chloroplast in cassava using phenol method;high resolution of 2⁃DE reference maps with good reproducibility were generated for proteomics;the comparison of the 2⁃DE gels for chloroplast of the 1,3,5 day under dark treatment found 186 differential protein spots;twenty⁃eight proteins were finally identi⁃fied by MALDI⁃TOF MS,which were involved in photosynthesis,carbon fixation,energy metabolism,proline metabolism,stress defense,and so on.This study represents a comprehensive dynamic protein profile and light⁃regulated network of SC8 cassava chloroplast under dark treatment.%以“华南8号”木薯叶绿体为研究材料，采用改进酚抽法提取总蛋白，通过一维和二维电泳，经Image Mas⁃ter分析，得到差异蛋白点进行质谱鉴定。结果表明：改进酚抽法可以从木薯叶绿体中提取高质量总蛋白；获得分离性好、分辨率及重复性较高的双向电泳图谱；比较黑暗条件下1，3，5 d木薯叶绿体的蛋白质表达谱，发现186个差异表达蛋白点；经MALDI－TOF MS质谱鉴定，获得28个差异蛋白，这些蛋白主要参与光合作用、碳固定、能量代谢、脯氨酸代谢、胁迫防御等代谢途径。本研究初步阐述了木薯叶绿体在黑暗条件下的蛋白质组变化，在蛋白质水平上鉴定出部分蛋白质的表达可能受光调控。