Abstract
Rhodococcus are gram-positive bacteria, which can exist in two different shapes rod and cocci. A number of studies have been done in the past on replication and stability of small plasmids in this bacterium; however, there are no reports on spatial localization and segregation of these plasmids. In the present study, a low copy number plasmid pDS3 containing pRC4 replicon was visualized in growing cells of Rhodococcus erythropolis PR4 (NBRC100887) using P1 parS-ParB-GFP system. Cells were initially cocci and then became rod shaped in exponential phase. Cocci cells were found to be non-replicating as evident by the presence of single fluorescence focus corresponding to the plasmid and diffuse fluorescence of DnaB-GFP. Rod shaped cells contained plasmid either present as one fluorescent focus observed at the cell center or two foci localized at quarter positions. The results suggest that the plasmid is replicated at the cell center and then it goes to quarter position. In order to observe the localization of plasmid with respect to nucleoid, plasmid segregation was also studied in filaments where it was found to be replicated at the cell center in a nucleoid free region. To the best of our knowledge, this is the first report on segregation of small plasmids in R. erythropolis.
Highlights
Rhodococcus are versatile and are known to have the ability to degrade a large variety of compounds [1]
In the present study we demonstrate the spatial localization of a small low copy number plasmid, pRC4 in R. erythropolis PR4
The system consists of a plasmid, which has ParB from plasmid P1 translationally fused to green fluorescence protein (GFP) under the control of an IPTG inducible lac promoter
Summary
Rhodococcus are versatile and are known to have the ability to degrade a large variety of compounds [1]. A number of studies have been done in the past on replication and stability of small plasmids in this bacterium; there are no reports on spatial localization and segregation of these plasmids. A low copy number plasmid pDS3 containing pRC4 replicon was visualized in growing cells of Rhodococcus erythropolis PR4 (NBRC100887) using P1 parS-ParB-GFP system.
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