Replacing Standard Reporters from Molecular Cloning Plasmids with Chromoproteins for Positive Clone Selection.

ABSTRACT: Cloning and expression plasmids are the workhorses of modern molecular biology. Despite the pathway paved by synthetic biology, laboratories around the globe still relay on standard cloning techniques using plasmids with reporter proteins for positive clone selection, such as β-galactosidase alpha peptide complementation for blue/white screening or ccdB, which encodes for a toxic DNA gyrase. These reporters, when interrupted, serve as a positive clone detection system. In the present report, we show that molecular cloning plasmids bearing the coding sequence for a 25.4 kDa protein, AmilCP, encoded by a 685 bp gene, that is well expressed in Escherichia coli, render blue-purple colonies. Using this reporter protein, we developed and tested a cloning system based on the constitutive expressi

SUBMITTER: Tafoya-Ramirez MD 

PROVIDER: S-EPMC6099721 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature