一个新型原核诱导表达系统的建立  

Establishment of a Novel Prokaryotic Inducible Expression System

作　　者：刘征[1] 高双成[2] 杨尚君[1] 孙宁 杜蕊[1] 赵彦宏[4] 

机构地区：[1]河北大学生命科学学院,保定071002 [2]河南科技大学农学院,洛阳471003 [3]保定师范附属学校,保定071000 [4]鲁东大学农学院,烟台264025

出　　处：《生物技术通报》2013年第8期145-149,共5页Biotechnology Bulletin

基　　金：教育部留学回国人员科研启动基金项目[(2011)1139号];河北大学引进人才项目(2010-185);山东省中青年科学家奖励基金项目(BS2009NY014);国家自然科学基金项目(30800682)

摘　　要：群体感应是细菌依赖于种群密度进行信息交流、协调群体行为并调控基因表达的一种方式。海洋发光弧菌Vibriofischeri利用该机制合成化学物质N-酰基-高丝氨酸内酯(AHLs),与LuxR蛋白结合,随后驱动下游生物荧光基因的高量表达。基于植物叶绿体起源于原始古细菌-蓝藻的假设以及叶绿体内的转录、翻译机制与原核生物高度近似,构建了luxR基因调控下游报告基因GFP表达的叶绿体遗传转化载体。将该载体转入大肠杆菌后,当无外加信号物质AHLs时,GFP不表达;而外加AHLs后,GFP大量表达,证明luxR和GFP基因的组合在大肠杆菌中是理想的化学诱导基因表达系统,可应用于大肠杆菌基因功能的研究。Quorum sensing is a term to describe bacterial communications,coordination of population behaviors and regulation of gene expressions depending on population density.Marine luminous bacterium Vibrio fischeri utilizes this mechanism to synthesize signal molecules N-Acyl homoserine lactones（AHLs）which serve as co-inducers with a transcriptional activator protein（LuxR）to regulate the expression of a set of target genes in an extremely high level.Based on the hypothesis that chloroplasts of higher plants were derived from cyanobacteria in ancient times and that the transcriptional and translational mechanisms of chloroplasts are highly similar to that of prokaryotes,we constructed that chloroplast transformation vector of the reporter gene GFP under the control of LuxR-dependent activation of the lux operon.The construct was then transformed into Escherichia coli.When no signal molecule of AHLs was added to the colonies,no GFP expression was detected.However,when AHLs were added,GFP expression was strong enough to be detected by naked eyes under ultraviolet lamp,demonstrating that the combination of theGFPgene and theluxoperon is an ideal chemical inducible gene expression system which can then be used for studies of gene function inEscherichia coli.

关 键 词：群体感应 LuxR基因 AHLs 化学诱导基因表达 叶绿体表达载体 

分 类 号：Q78[生物学—分子生物学]