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RESEARCH PRODUCT

Identification of SCP2165, a new SCP2-derived plasmid of Streptomyces coelicolor A3(2).

E. LecatMaria Grazia CusimanoSandra MarineoAnna GiardinaAnna Maria PugliaV. Di Caro

subject

Gel electrophoresisPlasmid preparationRecombination GeneticbiologyGene Transfer HorizontalSequence analysisStreptomycetaceaeStreptomyces coelicolorCloning vectorStreptomyces coelicolorbiology.organism_classificationApplied Microbiology and BiotechnologyStreptomycesMolecular biologyElectrophoresis Gel Pulsed-FieldPlasmidConjugative plasmids Pulsed-field gel electrophoresis SCP2 SCP2 derived plasmids Streptomyces coelicolorConjugation GeneticCrosses GeneticPlasmids

description

Aims:  Characterization of SCP2165, a plasmid identified in the Gram-positive bacterium Streptomyces coelicolor A3(2). Methods and Results:  Pulsed-field gel electrophoresis (PFGE) of mycelia of a S. coelicolor strain embedded in low melting agarose revealed the presence of a plasmid. Restriction enzyme mapping and sequence analysis of a 2·1 kb fragment revealed that this plasmid could be SCP2. SCP2 and its spontaneous derivative SCP2* are self-transmissible plasmids and have chromosome mobilizing ability (c.m.a.). SCP2* has a c. 1000-fold increased c.m.a. compared with SCP2. Interestingly the plasmid, named SCP2165, shows a c.m.a. from 5 × 10−2 to 1 × 10−1 which is 50–100-fold higher than that described for crosses involving SCP2*. Conclusions:  SCP2165 is a SCP2 derivative plasmid with the highest c.m.a. so far described for SCP2 derivative plasmids. PFGE, under conditions we used, seems to be a fast way to identify large circular plasmids in Streptomyces strains. Significance and Impact of the Study:  Further knowledge of the SCP2 family may allow the construction of improved SCP2-derived cloning vectors. SCP2165 could be a potential tool for conjugational transfer of gene clusters between different Streptomyces species.

10.1111/j.1472-765x.2005.01739.xhttps://pubmed.ncbi.nlm.nih.gov/16162143