Open AccessPlasmid copy number and stability determination in Clostridium perfringens transformants
Author(s) -
Allen Steven P.,
Blaschek Hans P.
Publication year - 1990
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.1574-6968.1990.tb03910.x
Subject(s) - plasmid , clostridium perfringens , biology , plasmid preparation , microbiology and biotechnology , dna , genetics , bacteria , pbr322
The copy number of the streptococcal plasmid pAMβ1 (26.5 kb), and its deletion derivatives, pVA1 (11 kb) and pVA677 (7.6 kb) contained in Clostridium perfringens 3624A transformants was determined by incorporation of [methyl‐ 3 ‐H]thymidine (4 μCi/ml) into chromosomal and plasmid DNA and sizing of the C. perfringens genome using transverse alternating field electrophoresis. Plasmids pAMβ1, pVA1, and pVA677 were found to be present at 1.0, 97, and 216 copies/cell, respectively, 10.2, 54, and 96% of the initial pAMβ1‐, pVA1‐ and pVA677‐containing transformants, respectively, remained resistant to erythromycin over 220 generations of growth. The results indicate a size‐dependent relationship between plasmid stability and plasmid copy number in C. perfringens .