摘要
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。
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.
出处
《核农学报》
CAS
CSCD
北大核心
2010年第5期926-931,共6页
Journal of Nuclear Agricultural Sciences
基金
教育部新世纪优秀人才基金(NCET-07-0251)
安徽省优秀青年人才基金(08040106803)
关键词
Aea-TMOF
植物表达载体
农杆菌转化
烟草
Aea-TMOF
plant expression vectors
transformation by Agrobacterium tunefacien
tobacco
