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当前位置: 首页 > 文章 > 苏云金杆菌Cry8Ea2蛋白的分离纯化及生物活性测定 华北农学报 2012,27 (6) 51-55
Position: Home > Articles > Separation,Purification and Insecticidal Activity of Cry8Ea2 Protein from Bacillus thuringiensis Acta Agriculturae Boreali-Sinica 2012,27 (6) 51-55

苏云金杆菌Cry8Ea2蛋白的分离纯化及生物活性测定

作  者：
王伟;郭巍;徐大庆;孙伟明;刘大群;李瑞军
单  位：
河北农业大学生命科学学院;河北农业大学植物保护学院
关键词：
苏云金杆菌;cry8;穿梭载体;蛋白纯化;生物活性测定
摘  要：
Cry8Ea2蛋白是一种新型苏云金杆菌杀虫晶体蛋白。通过穿梭载体pSXY422b将cry8Ea2导入Bt无晶体突变菌株中,获得工程菌HD8Ea2。Cry8Ea2蛋白在工程菌中获得表达。通过分子筛层析,成功获得纯化的130 kDa活性毒素蛋白。用改进的生物活性测定方法进行Cry8Ea2蛋白对华北大黑鳃金龟和暗黑鳃金龟一龄幼虫的杀虫活性测定,LC50分别为0.267 9μg/mL和0.071 9μg/mL。研究首次发现Cry8Ea2蛋白对华北大黑鳃金龟具有较高毒力,拓展了该蛋白的杀虫谱,为转基因工程提供了新资源。
译  名：
Separation,Purification and Insecticidal Activity of Cry8Ea2 Protein from Bacillus thuringiensis
作  者：
WANG Wei1,GUO Wei1,2,3,XU Da-qing1,SUN Wei-ming2,LIU Da-qun2,LI Rui-jun2(1.College of Life Sciences,Agricultural University of Hebei,Baoding 071001,China;2.College of Plant Protection, Agricultural University of Hebei,Biological Control Center of Plant Diseases and Plant Pests of Hebei Province, Baoding 071001,China;3.National Engineering Research Center for Agriculture in Northern Mountainous Areas,Baoding 071001,China)
关键词：
Bacillus thuringiensisis;cry8;Shuttle vector;Protein purification;Bioassays
摘  要：
Cry8Ea2 protein is a kind of insecticidal crystal protein of Bacillus thuringiensis.In this study,cry8Ea2 was cloned into E.coli-B.thuringiensist shuttle vector pSXY422b,resulting from recombinant pScry8Ea2,and pScry8Ea2 was transformed into the Bt,acrystalliferous mutant generating HD8Ea2 engineering strain.Cry8Ea2 protein was successfully expressed in HD8Ea2 stain.By anion exchange chromatography on molecular sieve chromatography,130 kDa of toxin protein Cry8Ea2 was purified.The activities of the Cry8Ea2 toxin were evaluated by improved bioassays,and the LC50 value to neonates of Holotrichia oblita and Holotrichia paralella was 0.267 9 μg/mL and 0.071 9 μg/mL,respectively.Insecticidal activity of Cry8Ea2 to Holotrichia oblita was firstly reported in this study,which will broaden the protein insecticidal spectrum and provide new resources for genetically modified project.

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