Open AccessSite‐specific integration of Streptomyces ΦC31 integrase‐based vectors in the chromosome of Rhodococcus equi
Author(s) -
Hong Yang,
Hondalus Mary K.
Publication year - 2008
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.1574-6968.2008.01298.x
Subject(s) - integrase , rhodococcus equi , plasmid , biology , streptomyces , genetics , electroporation , rhodococcus , bacterial conjugation , chromosome , transformation (genetics) , computational biology , microbiology and biotechnology , bacteria , virulence , dna , gene
Streptomyces ΦC31‐based site‐specific integration was used to transform the facultative intracellular pathogen Rhodococcus equi . The transformation efficiency of vectors incorporating the ΦC31 integrase and attP sites was comparable to that of replication plasmids using the same electroporation procedure. A single attB integration site was identified within an ORF encoding a pirin‐like protein, which deviates slightly from the consensus sequence of Streptomyces attB sites. Vector integration was stably maintained in the R. equi chromosome for as many as 100 generations during unselected passage in vitro . In addition, integration does not appear to affect the replication of bacteria inside macrophages. Finally, this integration system was also used to successfully complement an R. equi mutant.