Open AccessDevelopment of a gene knockout system for Ralstonia eutropha H16 based on the broad‐host‐range vector expressing a mobile group II intron
Author(s) -
Park Jong Myoung,
Jang YuSin,
Kim Tae Yong,
Lee Sang Yup
Publication year - 2010
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2010.02041.x
Subject(s) - ralstonia , cupriavidus necator , gene , bacteria , biology , group ii intron , intron , polyhydroxyalkanoates , genetics , computational biology , rna , rna splicing
Ralstonia eutropha H16 is a Gram‐negative lithoautotrophic bacterium and is one of the best biopolymer‐producing bacteria. It can grow to high cell densities either under lithoautotrophic or under heterotrophic conditions, which makes it suitable for a number of biotechnological applications. Also, R. eutropha H16 can degrade various aromatic compounds for environmental applications. The mobile group II intron can be used for the rapid and specific disruption of various bacterial genes by insertion into any desired target genes. Here, we applied the mobile group II intron to R. eutropha H16 and developed a markerless gene knockout system for R. eutropha : RalsTron. As a demonstration of the system, the phaC1 gene encoding polyhydroxyalkanoate synthase was successfully knocked out in R. eutropha H16. Furthermore, this knockout system would be useful for knocking out genes in other bacteria as well because it is based on a broad‐host‐range vector and the mobile group II intron that minimally depends on the bacterial hosts.