Fig. 2
From: Dissecting the contribution of Kup and KimA to Enterococcus faecalis potassium homeostasis
Intergenic region of the MntE and KimA genes in E. faecalis JH2-2. (a) Nucleotide sequence of intergenic region, the transcription and translation directions for both genes are indicated by a horizontal arrow, gray for MntE and orange for KimA. The putative Shine-Dalgarno (SD) sites are highlighted by the corresponding nucleotides, and the putative −10 hexamers for kimA are framed in violet. The presence of a putative rho-independent termination region is marked in bold. The predicted stems and loops are underlined. In italics are shown the primers that amplify the 574 bp fragment (b) On the left, schematic representation of the predicted rho-independent type terminator for MntE. In the center pKimA plasmid, derived from pTLGR, is schematized, constructed from a 574 bp fragment upstream of kimA in E. faecalis JH2-2. The black arrow indicates that the transcriptional fusion is directed towards the expression of the mCherry fluorescent protein. On the right, a schematic representation of the predicted rho-independent terminator for KimA. (c) Optical microscopy images of visible light with red fluorescence microscope (mCherry) of E. faecalis JH2-2 cells containing the pKimA vector are shown. Increase 100x. Scale bar 10 μm. (d) Activity of the transcriptional fusion of pKimA at an initial pH of 7 in M17 GA. Specific fluorescence was calculated as the ratio between the detected fluorescence, normalized to bacterial biomass estimated from the OD600 of the exponential growth culture. The average values of at least three independent experiments are presented, with error bars representing the standard deviations.
