Open AccessProduction of acidocin B, a bacteriocin of Lactobacillus acidophilus M46 is a plasmid‐encoded trait: Plasmid curing, genetic marking by in vivo plasmid integration, and gene transfer
Author(s) -
Vossen Jos M.B.M.,
Herwijnen Marcel H.M.,
Leer Rob J.,
Brink Bart,
Pouwels Peter H.,
Huis in `t Veld Jos H.J.
Publication year - 1994
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.1574-6968.1994.tb06724.x
Subject(s) - plasmid , puc19 , biology , bacteriocin , lactobacillus acidophilus , microbiology and biotechnology , plasmid preparation , lactobacillus plantarum , mutant , gene , bacteria , genetics , pbr322 , probiotic , antimicrobial , lactic acid
Plasmid‐curing studies suggest that acidocin B production is encoded by the 14‐kb plasmid pCV461 in Lactobacillus acidophilus M46. Loss of pCV461 from the original producer strain M46 did not coincide with loss of immunity to acidocin B. Bacteriocin activity determination after SDS‐PAGE showed that a substance of 2.4 kDa, absent in the culture supernatant of the mutant strain M46A2, lacking pCV461, represented acidocin B activity. In order to introduce a positive selection criterion, pCV461 was marked in vivo by the erythromycin resistance marker of pE194, present on pUC19 containing a 1.4‐kb Hin dIII fragment of pCV461, after plasmid integration. Introduction of this recombinant plasmid into the mutant strain M46A2 or Lactobacillus plantarum resulted in erythromycin‐resistant, acidocin B‐producing transformants, showing unambiguously that acidocin B is encoded by pCV461.