Wang Jian-Gang, Fan Chang-Sheng, Wu Yong-Qing, Jin Rui-Liang, Liu Dong-Xin, Shang Liang, Jiang Pei-Hong
Department of Microbiology, School of Life Sciences, Fudan University, Shanghai 200433, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Nov;35(11):993-7.
tyrR gene encodes a global regulatory protein (TyrR), which plays an important role in the transcriptional regulation of eight transcription units (including tyrR gene itself) whose protein products catalyze key steps in aromatic amino acid biosynthesis and/or transport. The aroP gene encodes an integral membrane protein (AroP) that transports aromatic amino acids through the cell membrane. The transcription of aroP was reported to be repressed by TyrR. In this work, aroP(p) (aroP gene carrying its own promoter), aroP (aroP gene without promoter) and tyrR genes were amplified by PCR from genomic DNA of E. coli K12 and introduced into E. coli WT5. The expression of aroP and tyrR were detected and the activities of AroP and TyrR were determined. The introduction of either aroP(p) or aroP elevated the strain's transport activity by 1.40 or 1.46-fold respectively. Transformant carrying tyrR gene showed an ATPase activity 1.69-fold compared with the control. When the genes were linked in tandem and co-expressed in a plasmid, the relative AroP transport activity of the strain harboring aroP(p) -tyrR (0.95) was significantly lower than that of aroP-tyrR (1.31). The results indicated that TyrR might be able to reduce the expression of aroP gene by binding with the aroP promoter region in E.coli.
tyrR基因编码一种全局调控蛋白(TyrR),它在八个转录单元(包括tyrR基因自身)的转录调控中发挥重要作用,这些转录单元的蛋白质产物催化芳香族氨基酸生物合成和/或转运中的关键步骤。aroP基因编码一种整合膜蛋白(AroP),它通过细胞膜转运芳香族氨基酸。据报道,aroP的转录受TyrR抑制。在这项工作中,通过PCR从大肠杆菌K12的基因组DNA中扩增出aroP(p)(携带自身启动子的aroP基因)、aroP(无启动子的aroP基因)和tyrR基因,并将其导入大肠杆菌WT5。检测了aroP和tyrR的表达,并测定了AroP和TyrR的活性。导入aroP(p)或aroP分别使菌株的转运活性提高了1.40倍或1.46倍。携带tyrR基因的转化体显示出的ATP酶活性是对照的1.69倍。当这些基因串联连接并在质粒中共表达时,携带aroP(p)-tyrR的菌株的相对AroP转运活性(0.95)显著低于aroP-tyrR(1.31)。结果表明,TyrR可能能够通过与大肠杆菌中的aroP启动子区域结合来降低aroP基因的表达。