title: Direct facile screening of recombinant DNA vector constructs.
abstract: Direct efficient facile screening of bacterial transformants with the goal of selecting, retrieving, and using recombinant DNA is exemplified by simple visual-based colorimetric inspections or fluorescent protein-based assays. We describe pRedScript, which introduces the constitutive expression of a very bright red fluorescent protein into transformants. On agar plates, red colonies are simply visualized in ambient white light in stark contrast to recombinant transformants that are white. In addition, the bright red fluorescence of the reporter protein can also be harnessed as a sensitive signal for screening bacterial promoters during the development of optimized fermentation conditions.
authors.affiliation: Division of Cancer Imaging Research, Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
authors.forename: Paul T
authors.initials: PT
authors.lastname: Winnard
authors.affiliation: Division of Cancer Imaging Research, Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
authors.forename: Rushi
authors.initials: R
authors.lastname: Challa
authors.affiliation: Division of Cancer Imaging Research, Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
authors.forename: Zaver M
authors.initials: ZM
authors.lastname: Bhujwalla
authors.affiliation: Division of Cancer Imaging Research, Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. Electronic address: vraman2@jhmi.edu.
authors.forename: Venu
authors.initials: V
authors.lastname: Raman
mesh_terms: D001483:Base Sequence; D004274:DNA, Recombinant / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D014170:Transformation, Genetic; D000097573:Red Fluorescent Protein
chemical_list: D004274:DNA, Recombinant; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: Recombinant DNA; Red color; Screening; mCherry