Premium
Construction of an R‐prime plasmid carrying the hup genes of Azorhizobium caulinodans ORS571 and its transfer to and stability in Rhizobium meliloti
Author(s) -
Miksch G.,
Lentzsch P.
Publication year - 1990
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.3620300310
Subject(s) - plasmid , biology , microbiology and biotechnology , gene , transfer dna , dna , hydrogenase , genetics , bacteria , transformation (genetics) , agrobacterium
An R‐prime plasmid (pJB3JI‐18) carrying a 10 kb region of Azorhizobium caulinodans ORS571 DNA which codes for uptake hydrogenase activity was isolated from a Km 8 derivative of R68.45. R‐prime plasmids were obtained by transferring R68.45 to ORS571, retransferring it to E. coli , and selecting transconjugants for hydrogenase activity by means of a colour test based on methylene blue. The R‐prime plasmid DNA was hybridized with a DNA region which contained the hup genes of Bradyrhiobium japonicum , indicating homology between the heterologous hup genes. Transfer of the R‐prime plasmid to different Hup − mutants restored the Hup + phenotype. When it was introduced to R. meliloti strains, it maintained its molecular weight and did not affect indigenous plasmids in two of three recipients. In the third recipient cointegrate formation and incompatibility with indigenous plasmids were found.