Zhao C Y, Yuan Z Q, Qin H M, Tian Y C
Plant Biotechnology Laboratory, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Sheng Wu Gong Cheng Xue Bao. 2001 May;17(3):273-7.
A synthetic Bt cry1Ac gene fussed with a secretary signal coding sequences at 5' end and a modified gna gene were used to construct a plant expression vector pBSGS1M+ and this vector was transferred into tobacco (Nicotiana tabacum L.) by Agrobacterium-mediated transformation method. Results of PCR, Southern blot and Slot blot analysis indicated that both the chimeric Bt cry1Ac and gna genes were integrated into the genomes of transformed plants. Western blot analysis indicated that at least the cry1Ac protein was produced in transgenic plants. Upon insect bioassay using cotton bollworm (Heliothis armigera Hubner), the mortality of insect larvae on 60% regenerated plants reached 100% in 5 days post infestation and the growth of the survived larvae was seriously inhibited; The results from insect bioassay with peach aphid (Myzus persicae) showed that the transgenic plants were aphid-resistant, evidenced by a 50%-60% reduction in aphid population density, even over 80% for some individual transgenic plants. These results reflect that the modification of the two insect resistant genes and construction of the expression vector are correct and could be valuable for later application in crop breeding for insect resistance.
一个在5'端与分泌信号编码序列融合的合成Bt cry1Ac基因和一个修饰的gna基因被用于构建植物表达载体pBSGS1M+,并且该载体通过农杆菌介导的转化方法被转入烟草(Nicotiana tabacum L.)。PCR、Southern杂交和狭缝杂交分析结果表明,嵌合的Bt cry1Ac和gna基因都已整合到转化植物的基因组中。Western杂交分析表明,转基因植物中至少产生了cry1Ac蛋白。在用棉铃虫(Heliothis armigera Hubner)进行昆虫生物测定时,60%再生植株上的昆虫幼虫在侵染后5天死亡率达到100%,存活幼虫的生长受到严重抑制;用桃蚜(Myzus persicae)进行昆虫生物测定的结果表明,转基因植物具有抗蚜性,蚜虫种群密度降低了50%-60%,一些单个转基因植物甚至超过80%。这些结果表明,两个抗虫基因的修饰和表达载体的构建是正确的,对以后在作物抗虫育种中的应用具有重要价值。