T vector bearing KillerRed protein marker for red/white cloning screening

Abstract

As a novel selection marker for DNA cloning, the genetically encoded photosensitizer KillerRed was used to achieve red/white cloning screening within the pZK18T T-vector system. KillerRed functioned without cofactors, inducers, or substrates. KillerRed-based red/white cloning screening was reliable in that bacteria containing DNA inserts that disrupt functional KillerRed expression form white colonies or red colonies as backgrounds. KillerRed simplifies assembly of customized vector and recombinant screening procedures. No special host or culture medium is required to amplify and prepare the vector in larger quantities. It makes high-throughput general-purpose cloning screening simpler, less expensive, and more effective.