Identification of biosynthetic gene clusters from metagenomic libraries using PPTase complementation in a Streptomyces host.

ABSTRACT: The majority of environmental bacteria are not readily cultured in the lab, leaving the natural products they make inaccessible using culture-dependent discovery methods. Cloning and heterologous expression of DNA extracted from environmental samples (environmental DNA, eDNA) provides a means of circumventing this discovery bottleneck. To facilitate the identification of clones containing biosynthetic gene clusters, we developed a model heterologous expression reporter strain Streptomyces albus::bpsA ΔPPTase. This strain carries a 4΄-phosphopantetheinyl transferase (PPTase)-dependent blue pigment synthase A gene, bpsA, in a PPTase deletion background. eDNA clones that express a functional PPTase restore production of the blue pigment, indigoidine. As PPTase genes often occur in biosyntheti

SUBMITTER: Bitok JK 

PROVIDER: S-EPMC5827622 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature