National Institute for Biotechnology and Genetic Engineering, P.O. Box 577, Jhang Road, Faisalabad, Pakistan.
Biotechnol Lett. 2013 Jun;35(6):969-74. doi: 10.1007/s10529-013-1153-2. Epub 2013 Feb 10.
A newly-synthesized cry2Ab gene was characterized in Nicotiana tabacum, before its further transformation in cotton. Synthetic cry2Ab gene was cloned in pGreen0029 and its expression was transiently analyzed at mRNA level through agroinfiltration in tobacco. The mRNA of cry2Ab was detected after 72 h agroinfiltration through PCR using total plant RNA. This construct was then transformed into N. tabacum through Agrobacterium. Insect bioassays were conducted on detached leaves using first instar Spodoptera exigua larvae; after 96 h significant insect mortality was recorded. This newly synthesized gene was effective in controlling S. exigua first instar larvae. It can be used in combinations with other Bt genes like cry1Ac for developing resistance against major insect pests of cotton and further widening the insect control spectrum.
新合成的 cry2Ab 基因在烟草中进行了特征描述，然后进一步在棉花中进行转化。合成的 cry2Ab 基因被克隆到 pGreen0029 中，并通过农杆菌浸润在烟草中进行了瞬时分析，在 mRNA 水平上进行了表达。通过使用总植物 RNA 进行 PCR，在 72 小时的农杆菌浸润后检测到 cry2Ab 的 mRNA。然后，该构建体通过根癌农杆菌转化到烟草中。使用初孵幼虫的第一龄斜纹夜蛾在离体叶片上进行昆虫生物测定；96 小时后记录到显著的昆虫死亡率。这个新合成的基因对控制第一龄斜纹夜蛾幼虫有效。它可以与其他 Bt 基因如 cry1Ac 结合使用，用于开发对棉花主要害虫的抗性，并进一步扩大昆虫控制谱。
