TA-GC cloning: A new simple and versatile technique for the directional cloning of PCR products for recombinant protein expression.

ABSTRACT: During the last few decades, the recombinant protein expression finds more and more applications. The cloning of protein-coding genes into expression vectors is required to be directional for proper expression, and versatile in order to facilitate gene insertion in multiple different vectors for expression tests. In this study, the TA-GC cloning method is proposed, as a new, simple and efficient method for the directional cloning of protein-coding genes in expression vectors. The presented method features several advantages over existing methods, which tend to be relatively more labour intensive, inflexible or expensive. The proposed method relies on the complementarity between single A- and G-overhangs of the protein-coding gene, obtained after a short incubation with T4 DNA polymerase, a

SUBMITTER: Niarchos A 

PROVIDER: S-EPMC5665511 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature