A strategy for building an amplified transcriptional switch to detect bacterial contamination of plants
A strategy for building an amplified transcriptional switch to detect bacterial contamination of plants
复制标题
DOI：
10.1007/s11103-011-9845-2
复制
发表时间：
2012-01-01
影响因子：
5.1
通讯作者：
Gurley, William B.
中科院分区：
文献类型：
--
作者：
Czarnecka, Eva;Verner, F. Lance;Gurley, William B.
我们已经设计并测试了一种转录自反馈回路，可以用来设计植物来感知细菌的存在。信号放大电路是基于对细菌鞭毛蛋白的生物开关而构建的。克隆了几种鞭毛蛋白和大肠杆菌诱导的拟南芥启动子，并利用基于鞭毛蛋白的肽在拟南芥和生菜原生质体中进行了瞬时表达试验。研究人员将这些基因作为报告基因的直接驱动因素，或作为转录自反馈回路的组成部分。拟南芥中木葡聚糖内转糖基化酶/水解酶18 （ATXTH18）启动子和细胞色素P450家族CYP82C3单加氧酶启动子作为生物开关作用良好。这些启动子被纳入我们的信号放大反馈回路系统。转录抑制因子的加入降低了基础表达，从而增加了信号检测的倍数放大，并微调了正自反馈回路调节。
We have designed and tested a transcriptional autofeedback loop that could be used to engineer plants to sense the presence of bacteria. The signal amplification circuit was built based on the biological switch responsive to the presence of bacterial flagellin. Several flagellin- and E. coli-inducible Arabidopsis promoters were cloned and tested in transient expression assays in Arabidopsis and lettuce protoplasts using a flagellin-based peptide. These were investigated either as direct drivers of a reporter gene, or as a component of a transcriptional autofeedback loop. Arabidopsis promoters from the xyloglucan endotransglucosylase/hydrolase 18 (ATXTH18) and cytochrome P450 family CYP82C3 monooxygenase worked well as biological switches. These promoters were incorporated into our feedback loop system for signal amplification. The inclusion of a transcriptional repressor reduced basal expression, thereby increasing fold-amplification of signal detection and fine-tuning the positive autofeedback loop regulation.