埃及伊蚊TMOF同源基因表达载体构建及烟草转化

胰蛋白酶调节抑制因子（Trypsin-modulating oostatic factor, TMOF）可以有效地抑制昆虫中肠胰蛋白酶的活性。本文为了研究埃及伊蚊TMOF同源物基因在转基因植物广谱抗虫中的作用，利用人工合成的Aea-TMOF同源物基因，以改造过的pCAMBIA1301质粒为基本载体，构建了由组成型CaMV35S启动子驱动Aea-TMOF基因的植物表达载体pCAMBIA1301+ Aea-TMOF。然后，采用冻融法转入农杆菌LBA4404菌株，通过叶盘法对烟草进行了遗传转化。经PCR和GUS染色检测，Aea-TMOF同源物基因已整合到烟草基因组中。

CONSTRUCTION OF PLANT EXPRESSION VECTORS CONTAINING Aedes aegypti TMOF ANALOGUE GENES, AND THEIR TRANSFORMATION TO TOBACCO

Trypsin-modulating oostatic factor (TMOF) can inhibit trypsin activity in insect midgut dramatically. To analyze inhibitory ability of Aedes aegypti TMOF analogues genes to insect, we reconstructed plant expression vectors containing Aedes aegypti TMOF analogue genes, and then transformed those into tobacco. The synthetic Aea-TMOF analogues were cloned into modified PCAMBIA1301 vector that contains constitutive promoter CaMV35S. The recombinant plasmids of PCAMBIA1301+Aea-TMOF were transferred into Agrobacterium tunefacien LBA4404 by freeze-thaw method; and then were used to infect tobacco leaf discs. Using PCR and GUS methods, we proved that the Aea-TMOF analogue genes were integrated into tobacco genome.