摘要
旨在为农业害虫防治提供更多的苏云金芽胞杆菌基因资源,从中国吉林市龙潭山土壤样品中分离得到野生菌株命名为Bt LTS-7,扫描电镜显示该菌株产生晶体形状为双锥体和正方体,聚丙烯酰胺凝胶电泳显示该菌株产生130 kD和71 kD的晶体蛋白,通过PCR鉴定出该菌株中含有cry2Ab和cry9Ea基因,并成功克隆到了这两个新基因,并被Bt国际命名委员会正式命名为cry2Ab28和cry9Ea9,将两个基因分别在大肠杆菌Rosetta(DE3)中表达,并进一步对其表达蛋白进行杀虫活性测定。结果显示,Cry2Ab28蛋白对棉铃虫(Helicoverpa armigera)初孵幼虫具有杀虫活性,LC50为32.45μg/mL。Cry9Ea9蛋白对小菜蛾初孵幼虫(Plutella xylostella)具有较高杀虫活性,LC50为0.77μg/mL。
In order to provide more gene resources for the insect pest biocontrol,Bacillus thuringiensis strain LTS-7 was isolated from soil samples at Longtan Mountain in Jilin of China.The strain formed the bipyramidal and square crystal which observed by scanning electron microscopy.SDS-PAGE analysis revealed molecular weight of insecticidal crystal protein was 130 kD and 71 kD in this strain.By use of PCR identification method,cry2Ab and cry9Ea genes were found in the Stain.The genes from strain Bt LTS-7 were identified and cloned,respectively.The two genes were designated as cry2Ab28 and cry9Ea9 by International Nomenclature Committee of Bt.Furthermore,two genes were respectively transformed into Escherichia coli Rosetta（DE3）strain and the toxicity of the two genes was evaluated.The bioassay results indicated that the Cry2Ab28 toxin had insecticidal activity against Helicoverpa armigera with LC50 32.45 μg/mL,and the Cry9Ea9 toxin had high insecticidal activity against Plutella xylostella with LC50 0.77 μg/mL.
出处
《生物技术通报》
CAS
CSCD
北大核心
2012年第10期100-105,共6页
Biotechnology Bulletin
基金
国家高技术研究发展计划课题(2011aa10a2O3)
转基因生物新品种培育科技重大专项(2009ZX08009-031B)
