柞蚕胚胎发育期蛋白质的2D-PAGE图谱分析

采用SDS-PAGE和2D-PAGE技术分析柞蚕胚胎期不同发育阶段的蛋白质组成和含量变化,为从蛋白质水平探讨柞蚕胚胎发育过程中的基因顺序表达模式积累基础信息。SDS-PAGE电泳结果显示:在柞蚕胚胎的发育过程中共分离到相对明显的32条蛋白带,包括分子质量约70、60、30 kD的高丰度蛋白条带,随着胚胎的发育,这些蛋白质含量逐渐减少,而40、15 kD左右的蛋白质含量逐渐增加,到孵化成蚁蚕(324 h)时,分子质量约70、60 kD的蛋白条带基本消失。进一步分析柞蚕胚胎发育不同时期蛋白质的2D-PAGE图谱:蛋白点数量随胚胎发育逐渐增多,胚胎发育12 h检测到370个蛋白点,胚胎发育300 h的蛋白点达到422个;胚胎发育132 h及276 h的蛋白点与胚胎发育早期36 h蛋白点的匹配率分别为52.4%和28.4%。此外,孵化蚁蚕总蛋白2D-PAGE图谱与胚胎期相比存在较大差异。研究结果提示,柞蚕胚胎发育前期,从胚胎形成期到附肢发生期(12～60 h),蛋白质种类相对较少,且变化不大;胚胎发育的气管形成期(228～276 h)开始,蛋白质种类变化剧烈,偏酸性蛋白质增加,蛋白点的匹配率下降。

2D-PAGE Image Analysis of Proteins from Developing Embryos of Antheraea pernyi

The present study analyzed variation of protein components and contents in Antheraea pernyi embryos of different developmental stages by SDS-PAGE and 2D-PAGE,so as to accumulate basic information on sequential expression of embryonic development genes in A.pernyi at the protein level.SDS-PAGE results showed that 32 major protein bands were isolated during the embryo development,including high abundance proteins with molecular weight of around 70,60 and 30 kD respectively.As the embryo development advanced,contents of these three proteins declined,while those of 40 and 15 kD proteins increased gradually.Proteins of 70 and 60 kD almost disappeared in newly-hatched larvae(324 h).Further analysis to 2D-PAGE images of embryonic proteins from different developmental stages showed that the protein spots increased gradually with embryo development.At 12 h of embryo development,there were 370 protein spots being detected.And at 300 h of embryo development,the number of protein spots reached 422.The matching rates of protein spots at 132 h and 276 h of embryo development with protein spots at 36 h of early embryo development was 52.4% and 28.4% respectively.More-over,2D-PAGE image of total proteins in newly-hatched larvae varied greatly with those at the embryonic stage.These results suggest that during the early stage of embryo development,i.e.from the embryo formation stage to appendage appearing stage(12～60 h),number of protein species were relatively low and did not change obviously;and after the trachea formation stage(228～276 h),protein species changed greatly,with increased number of acidic proteins and low matching rate of protein spots with previous stages.