Exploiting a precise design of universal synthetic modular regulatory elements to unlock the microbial natural products in Streptomyces.

ABSTRACT: There is a great demand for precisely quantitating the expression of genes of interest in synthetic and systems biotechnology as new and fascinating insights into the genetics of streptomycetes have come to light. Here, we developed, for the first time to our knowledge, a quantitative method based on flow cytometry and a superfolder green fluorescent protein (sfGFP) at single-cell resolution in Streptomyces. Single cells of filamentous bacteria were obtained by releasing the protoplasts from the mycelium, and the dead cells could be distinguished from the viable ones by propidium iodide (PI) staining. With this sophisticated quantitative method, some 200 native or synthetic promoters and 200 ribosomal binding sites (RBSs) were characterized in a high-throughput format. Furthermore, an insu

SUBMITTER: Bai C 

PROVIDER: S-EPMC4593075 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature