Xenbase Image ID: 153236

Figure 1: A construct containing mCherry and GFP coding regions separated by an in frame linker sequence allows the expression of a fusion protein in which the mCherry and GFP moieties fluoresce independently in bacteria. Top panel: Schematic representation of a plasmid construct for in-frame expression of mCherry and GFP (pmCherry-GFP). A sequence derived from pUC19 that contains multi-cloning sites (MCS) was inserted in between the mCherry and GFP coding regions under the control of the promoter of the bacterial lac operon (Plac). The inserted MCS sequence allows in frame translation of both mCherry and GFP separated by a linker peptide encoded by the MCS as shown. Bottom panel: When transfected into bacteria, pmCherry-GFP allows the expression of a fusion protein with both the mCherry and GFP moieties functioning independently. Bacteria were transformed with empty vector pUC19 (a), constructs expressing only mCherry (b) or GFP (c), or pmCherry-GFP (d), respectively. The transformed bacteria were plated and imaged in the bright field (A), or under a fluorescent microscope for red fluorescence (B) and green fluorescence (C), respectively. Red and green fluorescent imagines were merged in (D). Note that when transformed with pmCherry-GFP, both mCherry and GFP were functional, leading to a merged yellow image. Image published in: Fu L et al. (2016) Copyright © 2016, The Author(s). This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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