Open AccessIsolation of the replication region of an indigenous plasmid of Rhodobacter sphaeroides
Author(s) -
Barbé Jordi,
Castaño Maria,
Llagostera Montserrat,
Guerrero Ricardo
Publication year - 1986
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.1574-6968.1986.tb01762.x
Subject(s) - plasmid , bglii , biology , escherichia coli , rhodobacter sphaeroides , plasmid preparation , transposable element , genetics , restriction enzyme , restriction map , subcloning , origin of replication , microbiology and biotechnology , gene , pbr322 , bacteria , ecori , mutant
The isolation of the replication region of an indigenous plasmid of 42 kb of the phototrophic bacterium Rhodobacter sphaeroides is described. This plasmid was digested with the Bgl II restriction enzyme, ligated to the 2.7 Bgl II fragment of transposon Tn 10 , which contains the tet genes conferring tetracycline resistance, and the mixture was transformed into the Escherichia coli MC1061 strain. One of several chimeric plasmids harboring the replication region of the 42‐kb plasmid obtained by this process was named pUA33 and further characterized. Plasmid pUA33 is approx. 8.3 kb. A partial restriction map has been constructed. Plasmid pUA33 is stable in E. coli cells growing under non‐selective conditions and is non‐self‐transmissible. All these data suggest that the pUA33 plasmid may be a very useful tool for gene cloning in R. spheroides .