苏云金杆菌HBF-1菌株在土壤中毒蛋白含量的ELISA检测  

Detection of Bacillus thuringiensis Strain HBF-1 Protoxin in Soil by Enzyme-linked Immunosorbent Assay

作　　者：吴瑞华[1] 李国勋[2] 王容燕[3] 王金耀[3] 冯书亮[3] 

机构地区：[1]河北农业大学植物保护学院,保定071001 [2]莱阳农学院,青岛266109 [3]河北省农林科学院植保所,保定071000

出　　处：《中国生物防治》2007年第3期263-268,共6页Chinese Journal of Biological Control

基　　金：国家863计划(2006AA02Z189);河北省自然科学基金项目(C20050006420);河北省农科院青年基金项目(A06050101)

摘　　要：以苏云金芽孢杆菌HBF-1菌株中分离纯化的高特异性杀虫蛋白作为抗原,免疫新西兰白兔制备出多克隆抗体,其效价可达到16000。应用制备的特异性抗体建立了间接酶联免疫吸附测定法(ELISA),检测不同土壤样品中Bt毒蛋白含量。初步结果表明,该方法能够检测土壤中Bt毒蛋白的含量,因土壤理化特性及组成成分的不同,检测到的毒蛋白含量也略有不同,河沙中的蛋白检测量最低,只有12.53%;另一种沙壤混合物中检测量达到76.16%。With the SDS-PAGE preparation electrophoresis, high quality insecticidal protein was purified from a unique Bacillus thuringiensis serovar japonensis strain HBF-1, which is highly toxic to larvae of Anomala exoleta and A. corpulentathe. The protein was used as antigen and high immunity antiserum was got after several times of immuning to rabbits with titer reaching 1/16000. With the specific antiserum, a method for detecting the presence of the protoxin was developed. The Bt protoxin in different soil was determined by indirect ELISA. Results indicated that detection of the toxin was affected by soil texture. Detected rate in sandy soil was 12.53% only, and 76.16% in sandy loam soil.

关 键 词：HBF-1菌株 毒蛋白 多克隆抗体 间接ELISA 土壤 

分 类 号：S476.12[农业科学—农业昆虫与害虫防治]