Wu Jiahe, Luo Xiaoli, Wang Zhian, Tian Yingchuan, Liang Aihua, Sun Yi
Institute of Cotton Research and Biotechnology Research Centre, Shanxi Academy of Agricultural Sciences, Yuncheng, Taiyuan, PR China.
Biotechnol Lett. 2008 Mar;30(3):547-54. doi: 10.1007/s10529-007-9555-7. Epub 2007 Oct 16.
A synthetic scorpion Hector Insect Toxin (AaHIT) gene, under the control of a CaMV35S promoter, was cloned into cotton via Agrobacterium tumefaciens-mediated transformation. Southern blot analyses indicated that integration of the transgene varied from one to more than three estimated copies per genome; seven homozygous transgenic lines with one copy of the T-DNA insert were then selected by PCR and Southern blot analysis. AaHIT expression was from 0.02 to 0.43% of total soluble protein determined by western blot. These homozygous transgenic lines killed larvae of cotton bollworm (Heliothis armigera) by 44-98%. The AaHIT gene could used therefore an alternative to Bt toxin and proteinase inhibitor genes for producing transgenic cotton crops with effective control of bollworm.
在花椰菜花叶病毒35S启动子(CaMV35S)的控制下,通过根癌农杆菌介导的转化将合成蝎毒赫克托昆虫毒素(AaHIT)基因克隆到棉花中。Southern杂交分析表明,转基因整合到每个基因组中的拷贝数从1个到3个以上不等;然后通过PCR和Southern杂交分析筛选出7个T-DNA插入片段为单拷贝的纯合转基因株系。通过蛋白质免疫印迹法测定,AaHIT的表达量占总可溶性蛋白的0.02%至0.43%。这些纯合转基因株系对棉铃虫幼虫的致死率为44%-98%。因此,AaHIT基因可作为Bt毒素和蛋白酶抑制剂基因的替代品,用于培育有效防治棉铃虫的转基因棉花作物。