Wolk C P, Elhai J, Kuritz T, Holland D
MSU-DOE Plant Research Laboratory, Michigan State University, East Lansing 48824.
Mol Microbiol. 1993 Feb;7(3):441-5. doi: 10.1111/j.1365-2958.1993.tb01135.x.
The cyanobacterium Anabaena responds to nitrogen deprivation by producing heterocysts, cells specialized for nitrogen fixation, at well-spaced intervals along its filaments. The gene hepA, required for heterocyst maturation, is expressed in response to nitrogen deprivation, prior to visible differentiation. A spatial pattern of hepA expression indistinguishable from the eventual pattern of heterocysts was made visible by fusing the hepA promoter to luxAB, which encodes bacterial luciferase. Because the resulting signal did not greatly exceed instrumental background, T7 RNA polymerase was used to increase luminescence. The hepA promoter was fused to the gene for that polymerase, and a promoter recognized by that polymerase was fused to luxAB. Filaments containing these two fusions showed spaced luminescing cells many hours before differentiation became discernible morphologically.
蓝藻鱼腥藻通过在其丝状体上以规则间隔产生异形胞(专门用于固氮的细胞)来应对氮缺乏。异形胞成熟所需的hepA基因在氮缺乏时、在可见分化之前就会表达。通过将hepA启动子与编码细菌荧光素酶的luxAB融合,使与异形胞最终模式难以区分的hepA表达空间模式变得可见。由于产生的信号没有大大超过仪器背景,因此使用T7 RNA聚合酶来增强发光。将hepA启动子与该聚合酶的基因融合,并将该聚合酶识别的启动子与luxAB融合。含有这两种融合物的丝状体在形态上可辨别的分化出现许多小时之前就显示出间隔发光的细胞。