Open AccessThe genetic basis of plasmid tropism between C hlamydia trachomatis and C hlamydia muridarum
Author(s) -
Wang Yibing,
Cutcliffe Lesley T.,
Skilton Rachel J.,
Ramsey Kyle H.,
Thomson Nicholas R.,
Clarke Ian N.
Publication year - 2014
Publication title -
pathogens and disease
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.983
H-Index - 105
ISSN - 2049-632X
DOI - 10.1111/2049-632x.12175
Subject(s) - plasmid , chlamydia trachomatis , biology , transformation (genetics) , microbiology and biotechnology , virology , recombinant dna , replicon , tropism , genetics , gene , virus
The development of genetic transformation technology for C hlamydia trachomatis using its endogenous plasmid has recently been described. C hlamydia muridarum cannot be transformed by the C . trachomatis plasmid, indicating a barrier between chlamydial species. To determine which regions of the plasmid conferred the species specificity, we used the novel approach of transforming wild‐type C . muridarum carrying the endogenous plasmid pNigg and forced recombination with the C . trachomatis vector pGFP::SW2 which carries the complete C . trachomatis plasmid (pSW2). Penicillin and chloramphenicol‐resistant transformants expressing the green fluorescent protein were selected. Recovery of plasmids from these transformants showed they were recombinants. The differences between the pSW2 and pNigg allowed identification of the recombination breakpoints and showed that pGFP::SW2 had exchanged a ~ 1 kbp region with pNigg covering CDS 2. The recombinant plasmid (pSW2NiggCDS2) is maintained under antibiotic selection when transformed into plasmid‐cured C . muridarum . The ability to select for recombinants in C . muridarum shows that the barrier is not at transformation, but at the level of plasmid replication or maintenance. Our studies show that CDS 2, together with adjoining sequences, is the main determinant of plasmid tropism.