家蚕蛾触角蛋白的双向电泳分析  

Analysis of proteins in the antennae of Bombyx mori moths by two-dimensional electrophoresis

作　　者：王国宝[1] 陈玉华[1,2] 王举梅[1] 卫正国[1,2] 许雅香[1,2] 李兵[1,2] 沈卫德[1,2] 

机构地区：[1]现代丝绸国家工程实验室,江苏苏州215123 [2]苏州大学蚕桑研究所,江苏苏州215123

出　　处：《昆虫学报》2011年第5期589-595,共7页Acta Entomologica Sinica

基　　金：国家现代农业产业技术体系-家蚕生理生态岗(nycytx-27-gw307)

摘　　要：为探讨家蚕Bombyxmori蛾触角发生发育、结构与功能的分子基础和调控机制,我们采用双向电泳结合基质辅助质量飞行时间质谱（MALDI-TOF/MS）技术初步探讨了家蚕蛾触角蛋白及其雌雄表达差异。采用ImageMast 6.0软件分析电泳图谱,在家蚕蛾触角中检测到约550个蛋白点,主要集中在分子量14~70 kD,等电点4~8之间。从雌雄蛾触角电泳图谱中分别检测到419和489个蛋白点,其中雌雄匹配蛋白点有326对,匹配率为71.81%。雌雄间表达差异点有34个,雌雄分别所特有的特异蛋白点分别为9和20个。对特异和差异点进行MALDI-TOF/MS鉴定获得其中5种蛋白——成虫原基生长因子（imaginal disk growth factor）、表皮蛋白RR-1基序15（cuticular protein RR-1 motif15）、硫醇过氧化还原酶（thiol peroxiredoxin）、空泡ATP酶B亚基（vacuolar ATPase B subunit）以及gasp前体（gaspprecursor）,它们在雄蛾触角中表达量都比雌蛾高。这为家蚕触角蛋白的进一步研究提供了基本信息。To investigate the molecular basis and regulatory mechanism of antennal development,structure and function of Bombyx mori,high resolution and two-dimensional polyacrylamide gel electrophoresis followed by matrix-assisted laser desorption ionization and time of flight mass spectrometry（MALDI-TOF/MS） was applied to analyze the total proteins in antennae and their expression difference between male and female moths.The ImageMaster 6.0 software was used to analyze the two-dimensional gel electrophoresis profiles.Approximately a total of 550 protein spots were detected in the antenna of silk moth,most of which were arranged from 14 to 70 kD with pI value 4-8.In the two-dimensional gel electrophoresis profiles of male and female antenna,419 and 489 protein spots were detected,respectively,and the number of matched protein spots between male and female was 326.The matching rate was 71.81%.There were 34 protein spots which have different gray value but were distributed in both gel electrophoresis profiles of male and female antenna.Meanwhile,9 and 20 specific protein spots were detected in gel electrophoresis profiles of female and male,respectively.Sixty-three protein spots were identified by MALDI-TOF/MS and five proteins were obtained,including imaginal disk growth factor,cuticular protein RR-1 motif 15,thiol peroxiredoxin,vacuolar ATPase B subunit and gasp precursor.These proteins are involved in cell proliferation,epidermis formation,the removal of H2O2 in vivo,building up of a H＋ gradient,etc.These five proteins were expressed differentially between female and male antenna,and all had higher expression levels in male antenna than in female antenna.The results provide some basic data for the further research of antenna proteins of the silk moth.

关 键 词：家蚕 触角 蛋白质组学 双向电泳 MALDI-TOF/MS 质谱分析 

分 类 号：Q965[生物学—昆虫学]