Tn10介导的Bit cryIVA基因在荧光假单胞菌染色体中的整合及表达

苏云金杆菌以色列亚种的杀虫晶体蛋白基因cryIVA被亚克隆到自杀型转座子质粒载体pLOF／Km的TN10中，构建了转座子质粒PLF97A。通过电转化／转座作用，cryIVA随Tn10转座并整合到荧光假单胞菌FP．DE2染色体中，构建了工程菌株FP．DE202。Southernblotting验证了cryIVA在FP．DE202中整合在不同的位点。Westernblotting证明了cryIVA在F．P．DE202中得到了表达，其产物对双翅目害虫韭菜迟眼蕈蚊的3龄幼虫有较强的毒杀效果。

Integration via Tn10 and Expression of Bit cryIVA Gene in the Chromosome DNA of Flourescent Pseudomonas Bacterial Strains

cryIVA gene of the Bacillus thuringiensis subsp. israelensis was subeloned into the Tn10 of a suicide transposon plashed vector and a transposon plasmid pLF97A carrying cryIVA was constructed. By eletroporation and transposihon, cryIVA gene integratod into chromosome DNA of F. P. DE2. Thus the genetic engineered bacterial strain, F.P.DE202, was constructed. Southern blotting revealed that thes integration was happened in different loci of the chromosome. Western blotting demonstrated that cryIVA was expressed in the engineered strain, and that the expressed product possesed the peshcidal ability against the third-instar larva of Bradysia odoriphaga.