This study was conducted to build a recombinant strain with highly insecticidal activity and a wide host range by using the crylAc and p74 gene. Firstly, the p74 gene was amplified from the genosome ofAutographa calif...This study was conducted to build a recombinant strain with highly insecticidal activity and a wide host range by using the crylAc and p74 gene. Firstly, the p74 gene was amplified from the genosome ofAutographa californica multicapsid nucleopolyhedrovirus. The crylAc gene and the terminator gene of crylAc, named crylAct, were amplified from the plasmid of Bt 4.0718 strain. Three T vectors, named pTp74, pT1Ac, and pT1Act which held the aimed gene p74, cry1Ac, and crylAct, respectively, and two middle vectors, named pTp74Act and pTIAcp74 which held the aimed fusion gene p74-crylAct and cry lAc-p74, respectively, were built by using pMDI 8-T. Then pTiAcp74 and the shuttle plasmid were digested and linked and an expressing-vector pH1Acp74 was built. Finally, pH1Acp74 was transformed into the acrystalliferous strain XBU001 and the aimed recombinant strain XBU-H1Acp74 was obtained. The expression of Bt transformant XBU-H1Acp74 was analyzed by SDS-PAGE which showed XBU-H1Acp74 could produce 130 kDa CrylAc protein and 50 kDa P74 protein. The insecticidal activity of transformant against Spodoptera exigua was evaluated compared with the contrast strains HTX-42 （only crylAc gene was transformed into XBU001） after autolysis. The LCs0 of HTX-42 was higher than that of the XBU-H IAcp74＇s, which implied that P74 could increase the efficacy and range of Bt Cry toxins in insect control. The fusion gene of crylAc and p74 were constructed successfully which will be served as the foundation lbr constructing the fusion genes of Bt cry gene and other foreign genes.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.31272100,31372199)the Natural Science Foundation of Shanxi Province,China(No.2014011038-1)+1 种基金the National High Technology Research and Development Program of China(863 Program,No.2012AA020809)the Program for the Top Young Academic Leaders of Higher Learning Institutions of Shanxi Province,China
基金supported by the National Natural Science Foundation of China (30570050, 30670052)the National High Technology R&D Program of China (863Program, 2006AA02Z187)+1 种基金the National Ph D Programs Foundation of China (200-60542006)the Natural Science Foundation of Hunan Province, China(06JJ50062, 06JJ2009).
文摘This study was conducted to build a recombinant strain with highly insecticidal activity and a wide host range by using the crylAc and p74 gene. Firstly, the p74 gene was amplified from the genosome ofAutographa californica multicapsid nucleopolyhedrovirus. The crylAc gene and the terminator gene of crylAc, named crylAct, were amplified from the plasmid of Bt 4.0718 strain. Three T vectors, named pTp74, pT1Ac, and pT1Act which held the aimed gene p74, cry1Ac, and crylAct, respectively, and two middle vectors, named pTp74Act and pTIAcp74 which held the aimed fusion gene p74-crylAct and cry lAc-p74, respectively, were built by using pMDI 8-T. Then pTiAcp74 and the shuttle plasmid were digested and linked and an expressing-vector pH1Acp74 was built. Finally, pH1Acp74 was transformed into the acrystalliferous strain XBU001 and the aimed recombinant strain XBU-H1Acp74 was obtained. The expression of Bt transformant XBU-H1Acp74 was analyzed by SDS-PAGE which showed XBU-H1Acp74 could produce 130 kDa CrylAc protein and 50 kDa P74 protein. The insecticidal activity of transformant against Spodoptera exigua was evaluated compared with the contrast strains HTX-42 （only crylAc gene was transformed into XBU001） after autolysis. The LCs0 of HTX-42 was higher than that of the XBU-H IAcp74＇s, which implied that P74 could increase the efficacy and range of Bt Cry toxins in insect control. The fusion gene of crylAc and p74 were constructed successfully which will be served as the foundation lbr constructing the fusion genes of Bt cry gene and other foreign genes.
