【作者】 洪华珠； 刘凯于； 郑丙莲； 蒋才富； 黄薇； 彭建新；

【Author】 Huazhu Hong,Kaiyu Liu,Binglian Zheng,Caifu Jiang,Wei Huang and Jianxin Peng~* (Key Laboratory of pesticide and Chemical Biology,Minist ry of Education, Huazhong Normal University,Wuhan 430079)

【机构】 华中师范大学农药与化学生物学教育部重点实验室,华中师范大学昆虫学研究所；

【摘要】 苏云金芽孢杆菌(Bt)是一种安全、有效、环境友好型生物杀虫剂,随之广泛应用,害虫也对BT的毒素产生了高水平的抗性,因此进一步了解抗性发生的分子机制在Bt的应用中十分重要。本文对选育的抗Bt Cry1Ac毒素的细胞进行一些分子生物学比较研究。发现敏感细胞的膜蛋白制备物中存在两个与Bt Cry 1Ac毒素特异性结合的靶位点,分子量分别为37KD和34 KD;而抗性细胞膜制备物中只有34KD处的蛋白是与Cry1Ac毒素特异性结合的靶位点。并且两种细胞的在抗性细胞的2-DE总蛋白图谱上检测到的平均蛋白质点数可达到707±25个,在敏感细胞的总蛋白2-DE图谱上检测到的平均蛋白质点数可达到637±19个,其中分辨率高重复性较良好的显著差异点有10多个。对其中一些差异斑点测定了肽质指纹图谱,经数据库查询,初步鉴定差异蛋白质为一些与胞质外周蛋白,脂多糖生物合成蛋白和免疫球蛋白重链等具同源性的蛋白质。在敏感和抗性两种细胞的2-DE总膜蛋白图谱上检测到的平均蛋白质点数可达到260多个,其中显著差异点有10余个。更多还原

【Abstract】 Bacillus thuringiensis are being widely used effectively for control of agricul- turally and biomedically detrimental pests because the bio-insecticides are environmentally friendly.However,the pests might develop high-level resistance to Bt toxins rapidly.Further understanding of the mechanism is fundamental in sustaining the use of Cry proteins in integrated pest management.Here,the authors had selected the resistant cultured insect cells to Bt Cry toxin instead of resistant insects,and studied on the molecular-biological characterization of resistance.The results were shown as followed: Primary results have shown that the toxin binding to the membrane of resistant cells was less than that of susceptible cells,and a protein binding to CrylAc was missed in the resistant cell membrane.There was gene differential expression between the resistant and susceptible cells.The proteome and sub-proteome difference were analyzed between the resistant and susceptible cells using 2-DE and peptide mass fingerprint technology.707 + 25 spots were detected on the resistance cell proteomic 2-DE map,and 637±19 on the that of susceptible cells with Melanie Viewer E software.Differential membrane proteins also were detected between the resistant and susceptible cells on 2-DE images.Some differential spots were measured using the peptide mass fingerprint technology,and searched in the database.更多还原