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ResultsMcaSRNAinductionduringthetransitionintostationaryphaseTheE.coliMcaS(IS061/IsrA... Results
McaS RNA induction during the transition into
stationary phase
The E. coli McaS (IS061/IsrA) RNA was previously pre-dicted in a search for orphan promoter and terminator
sequences and suggested to be ~ 158 nt in length (Chen
et al ., 2002). The sRNA is encoded in an intergenic region
between the gene encoding a LysR-type transcription
factor, AbgR, a regulator of the divergently encoded
p-aminobenzoyl glutamate catabolism operon abgABT
(Hussein et al ., 1998), and a gene encoding a small MutS-related protein, YdaL (Gui et al ., 2011) (Fig. 1A and B).
Based on sequence conservation, McaS is present in other
E. coli and Shigella strains, but is absent from most enteric
bacteria such asSalmonella (Zhang et al ., 2003) (data not
shown).
To validate McaS expression, we probed total RNA
isolated from wild-type MG1655 grown in Luria–Bertani
(LB) media over a period of 30 h (Fig. 1C and Fig. S1). We
detected a single predominant band of ~ 95 nt that was low
during exponential phase and increased during the transi-tion from exponential to stationary phase. Maximal McaS
expression was reached around~ 4–7 h of growth (OD600
~ 2.5–4.0) with levels decreasing as the cells entered late
stationary phase at ~ 30 h. A strain carrying 227 nt of the
McaS promoter fused to alacZ reporter [P
mcaS(227 )–lacZ]
also showed an increase in b-galactosidase activity during
the transition into stationary phase (Fig. S2). However, the
activity did not decrease later in stationary phase, most
likely due to the stability of the b-galactosidase protein.
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结果
McaS RNA诱导在过渡到
固定相
大肠杆菌的平均血流速度(IS061 /伊斯拉)RNA曾pre-dicted在搜索孤儿发起人和终结者
序列和建议~ 158元的长度(陈
et al。,2002)。可溶rna编码的在一个基因间区
基因编码一个LysR-type之间的转录
因素,AbgR,监管者的divergently编码
p-aminobenzoyl谷氨酸分解代谢abgABT操纵子
(侯赛因et al。,1998),和一个基因编码一个小杂种狗相关的蛋白质,YdaL(Gui et al。,2011)(无花果。1 a和B)。
基于序列保护,McaS存在于其他
大肠杆菌、志贺氏杆菌菌株,但缺席大部分肠
细菌能够假借它们污染鸡蛋(Zhang et al等。,2003)(数据没有
如图所示)。
验证McaS表达式,我们探索总RNA
孤立于野生型生长在Luria-Bertani MG1655
(磅)媒体在一段30 h(无花果。1 c和无花果。S1)。我们
检测到一个主要的群~ 95元,很低
在指数期,增加了在中的优化指数,从固定相。最大血流
表达了~ 4 - 7 h的周围(OD600增长
~ 2.5 - -4.0)与水平降低的细胞进入后期
固定相在~ 30 h。一个应变携带227元的
启动子融合到alacZ McaS记者(P
平均血流速度(227)-lacZ]
也显示增加b牛乳糖活动的过程
转变成固定相(无花果。S2)。然而,
活动没有减少后在固定相,大多数
可能由于稳定性的b牛乳糖蛋白质。
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