Zhang X D, Letham D S, Zhang R, Higgins T J
CSIRO Division of Plant Industry, Canberra, Australia.
Transgenic Res. 1996 Jan;5(1):57-65. doi: 10.1007/BF01979922.
The isopentenyl transferase gene (ipt) from Agrobacterium tumefaciens was isolated and introduced, via a disarmed binary vector, into tobacco using the Agrobacterium tumefaciens-mediated gene transfer system. The expression of the ipt gene was monitored by RNA hybridization, western blotting and cytokinin analysis. The addition of auxin to the media rapidly reduced the level of cytokinins in the transgenic tissues and this was associated with a reduction in IPT mRNA and protein levels. It is concluded that the hormone auxin can regulate expression of a gene involved in biosynthesis of the second hormone cytokinin. Although exogenous benzyladenine did not directly affect ipt gene expression, it did antagonize the effect of auxin on levels of cytokinins and IPT mRNA and protein.
从根癌农杆菌中分离出异戊烯基转移酶基因(ipt),并通过无毒性双元载体,利用根癌农杆菌介导的基因转移系统将其导入烟草。通过RNA杂交、蛋白质免疫印迹和细胞分裂素分析对ipt基因的表达进行监测。向培养基中添加生长素会迅速降低转基因组织中细胞分裂素的水平,这与IPT mRNA和蛋白质水平的降低有关。得出的结论是,激素生长素可以调节参与第二种激素细胞分裂素生物合成的基因的表达。虽然外源苄基腺嘌呤没有直接影响ipt基因的表达,但它确实拮抗了生长素对细胞分裂素水平以及IPT mRNA和蛋白质的影响。