Liu J, Song F, Zhang J, Liu R, He K, Tan J, Huang D
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100094, P.R. China.
Lett Appl Microbiol. 2007 Oct;45(4):432-8. doi: 10.1111/j.1472-765X.2007.02217.x. Epub 2007 Sep 14.
To search for novel Vip3A proteins for controlling insect pests.
A pair of universal primers was designed based on the conserved regions of five vip3A genes. Amplified products were digested with the HindIII and EcoR enzymes so as to confirm different restriction fragment length polymorphism (RFLP) patterns used to identify vip3A-type genes. The vip3A gene types of 606 Bacillus thuringiensis strains were screened and three patterns of RFLP were successfully identified. Two novel vip3A genes were found and one of these, vip3Aa19, was further characterized and its product was confirmed toxic to Spodoptera exigua, Helicoverpa armigera and Plutella xylostella larvae. Partial sequences of another novel vip3A-type gene were obtained that shared 83% homology with that of the vip3Af1 gene.
A polymerase chain reaction (PCR)-RFLP system we developed could be used for identifying novel vip3A-genes from B. thuringiensis strains. A novel Vip3A protein was found to have a broader insecticidal spectrum.
The reported method is a powerful tool to find novel Vip3A proteins from large-scale B. thuringiensis strains. The novel Vip3A protein may be used to control insect pests or resistant insect pests by constructing genetically engineered strains or transgenic plants.
寻找用于控制害虫的新型Vip3A蛋白。
基于5个vip3A基因的保守区域设计了一对通用引物。用HindIII和EcoR酶消化扩增产物，以确认用于鉴定vip3A类型基因的不同限制性片段长度多态性（RFLP）模式。筛选了606株苏云金芽孢杆菌菌株的vip3A基因类型，成功鉴定出3种RFLP模式。发现了两个新型vip3A基因，其中一个vip3Aa19进一步进行了特性分析，其产物被证实对甜菜夜蛾、棉铃虫和小菜蛾幼虫有毒性。获得了另一个新型vip3A类型基因的部分序列，与vip3Af1基因的部分序列具有83%的同源性。
我们开发的聚合酶链反应（PCR）-RFLP系统可用于从苏云金芽孢杆菌菌株中鉴定新型vip3A基因。发现一种新型Vip3A蛋白具有更广泛的杀虫谱。
所报道的方法是从大规模苏云金芽孢杆菌菌株中寻找新型Vip3A蛋白的有力工具。新型Vip3A蛋白可通过构建基因工程菌株或转基因植物用于控制害虫或抗性害虫。
