Open AccessMultifunctional SEVA shuttle vectors for actinomycetes and Gram‐negative bacteria
Author(s) -
GarcíaGutiérrez Coral,
Aparicio Tomás,
TorresSánchez Lucía,
MartínezGarcía Esteban,
Lorenzo Víctor,
Villar Claudio J.,
Lombó Felipe
Publication year - 2020
Publication title -
microbiologyopen
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.881
H-Index - 36
ISSN - 2045-8827
DOI - 10.1002/mbo3.1024
Subject(s) - biology , streptomyces , shuttle vector , plasmid , bacteria , genetics , computational biology , microbiology and biotechnology , vector (molecular biology) , gene , recombinant dna
Actinomycetales , such as the genus Streptomyces , are well‐known cell factories employed to produce a wide variety of secondary metabolites for industrial use. However, not only is the genetic engineering of Streptomyces more complicated and tedious than other model laboratory species, such as Escherichia coli, there is also a considerable lack of genetic tools, hindering its adoption as a common chassis for synthetic biology . In this work, 23 novel shuttle vectors are presented that follow the canonical SEVA (Standard European Vector Architecture) common architecture with the goal of increasing the genetic toolbox repertoire for Streptomyces and other actinomycetes. The ORI module of these plasmids is composed of the combination of two origins of replication, one for Gram‐negative bacteria and the other for Streptomyces , a Gram‐positive bacteria. Origins of replication have been included in the collection for integrative, low‐copy number, and medium‐to‐high‐copy number vectors for Streptomyces. Also, a new selection marker has been developed that confers resistance to apramycin. The functionality of these plasmids was tested via the heterologous expression of GFP and the heterologous production of the plant flavonoid apigenin in Streptomyces albus J1074, with successful results in both cases, therefore expanding the current repertoire of genetic manipulation tools in Streptomyces species.