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. 2018 Nov 16;9:2674. doi: 10.3389/fmicb.2018.02674

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Copyright © 2018 Chandrashekhar, Srivastava, Hwang, Jeon, Ryu and Rajashekara.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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(A) Genetic organization of tlp2. The tlp2 gene (cjj81176_180) is located upstream of the phoX gene (Cjj81176_181) which encodes the alkaline phosphatase (PhoX) enzyme. The tlp2 and phoX genes are separated by a 135 bp intergenic region. (B) Reverse Transcriptase overlapping PCR showing co-transcription of tlp2 and phoX. Intergenic region was amplified with primer pair P1 F (forward) and R (reverse) in WT strain grown in MEM-α. Primers for the tlp2 (P2 F and R) and phoX (P3 F and R) genes were included as control regions as well (data not shown). (C) The P_int-lacZ fusion showed no β-galactosidase activity compared to the P_tlp2–lacZ fusion in the WT strain. Determination of the transcriptional start site for tlp2 (D) and phoX (E) by a primer extension assay. Only one transcriptional start site is seen upstream to tlp2, designated TS (Transcriptional Start) indicated with an arrowhead on the right and by the ^∗ in the sequence. No transcriptional start site was found in the region upstream to phoX (F): The −10 and −35 elements of the P_tlp2 are underlined and the ribosomal binding site is indicated as RBS.