Regulation of aroP expression by tyrR gene in Escherichia coli.

Jian-Gang Wang, Chang-Sheng Fan, Yong-Qing Wu, Rui-Liang Jin, Dong-Xin Liu, Liang Shang, Pei-Hong Jiang
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Abstract

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.

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tyrR基因对大肠杆菌aroP表达的调控。
tyrR基因编码一个全局调控蛋白(global regulatory protein, tyrR),该蛋白在包括tyrR基因本身在内的8个转录单位的转录调控中起重要作用,其蛋白产物催化芳香氨基酸生物合成和/或转运的关键步骤。aroP基因编码一种完整的膜蛋白(aroP),该蛋白通过细胞膜运输芳香氨基酸。据报道aroP的转录被TyrR抑制。本研究从大肠杆菌K12基因组DNA中扩增出aroP(p)(携带自身启动子的aroP基因)、aroP(不带启动子的aroP基因)和tyrR基因,并将其导入大肠杆菌WT5。检测aroP和tyrR的表达,测定aroP和tyrR的活性。aroP(p)和aroP的引入分别使菌株的转运活性提高了1.40倍和1.46倍。携带tyrR基因的转化体的atp酶活性是对照的1.69倍。当基因串联在质粒中共表达时,AroP (p) -tyrR菌株的相对AroP转运活性(0.95)显著低于AroP -tyrR菌株(1.31)。结果表明,TyrR可能通过与大肠杆菌aroP启动子区结合来降低aroP基因的表达。
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