Site-specific integration of Streptomyces PhiC31 integrase-based vectors in the chromosome of Rhodococcus equi |
| |
Authors: | Hong Yang Hondalus Mary K |
| |
Affiliation: | Department of Infectious Diseases, University of Georgia, Athens, GA, USA. |
| |
Abstract: | StreptomycesPhiC31-based site-specific integration was used to transform the facultative intracellular pathogen Rhodococcus equi. The transformation efficiency of vectors incorporating the PhiC31 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. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|