苏云金芽胞杆菌营养期杀虫蛋白Vip与Cry蛋白的协同作用

将构建的营养期杀虫蛋白基因vip3表达质粒pBMB2328和含杀虫晶体蛋白基因(crylAc10或crylCa)质粒同时电转化无质粒突变株BMB171并双抗筛选。经PCR特异引物扩增验证，分别得到含crylAc10和vip83、crylCa和vip83的双基因重组菌BMB2830-171和BMB2882-171。用单基因重组菌作对照，分别测定了营养期杀虫蛋白Vip83与杀虫晶体蛋白CrylAc10和Cry1Ca两组蛋白对3种重要鳞翅目害虫毒力。经统计分析，结果表明两组杀虫蛋白Vip83与CrylAc10和Vip83与CrylCa之间对棉铃虫均存在拮抗作用，对甜菜夜蛾协同作用不明显；但对小菜蛾前协同作用不明显，而后则有增效作用，其共毒系数为32．6。双基因的遗传稳定性检测表明这种正负协同关系具有一定的分子遗传稳定性，可为高效广谱工程菌的构建提供依据。

THE SYNERGISM OF VEGETATIVE INSECTICIDAL PROTEIN VIP AND CRY PROTEIN FROM BACILLUS THURINGIENSIS

In this paper, vegetative( vip83 ) and crystal(cry1Ac10 and cry1Ca) insecticidal protein genes from Bacillus thuri ngiensis were simultaneously electrospored into the plasmid-free strain BMB17 1. By the means of the specific P CR detection, the recombinant strains BMB2830-171 contained cry 1Ac10 and vip83, and BMB2 882-171 had cry1Ca and vip83 , were obtained respectively. Under the control of r ecombinant strains with one gene, bioassay of the synergism between vegetative V ip83 and crystal Cry1Ac10( or Cry1Ca )insecticidal proteins to three important Lepidopteran pests were done. The results, by analysis of statistic bio-so ft, showed that the synergia relation of vegetative Vip83 and crystal Cry1Ac10 i nsecticidal protein toxic to Heliothis armigera wascounteracted, while Plu tella xylostella and Spodotera exigua unobservable. There was no synergis tic action between Vip83 and Cry1Ca insecticidal proteins with Spodotera exigu a as tested insect. Bu t their cooperation to Heliothis armigera was minus, and the counterpart to Plutella xylostella plus, whose cotoxicity factor is 32.6. The experiment of bi-g ene genetic stability also suggested that the synergia effection had certain molecu lar genetic stability in the same cell. This performance can be contributed to construct high-effect and wide-spectrum engineered strain.