Site-specific degradation of Streptomyces lividans DNA during electrophoresis in buffers contaminated with ferrous iron.
Streptomyces lividans DNA contains a modification which makes it susceptible to double-strand cleavage during electrophoresis in buffers contaminated with ferrous iron (which may be present in some batches of EDTA). The cleavage of the DNA is site-specific and the average fragment size resulting from limit digestion of total S. lividans DNA is about 6kb. DNA from Streptomyces coelicolor A3(2) and several other Streptomyces strains, and from E. coli, is not cleaved under the same conditions. A S. lividans mutant has been isolated which lacks the DNA modification. We suspect that many reports of "poor" preparations of S. lividans plasmids may be due to the above effect.
Zhou
X
X
John Innes Institute, Norwich, UK.
Deng
Z
Z
Firmin
J L
JL
Hopwood
D A
DA
Kieser
T
T
D002021:Buffers; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D005822:Genetic Vectors; D007501:Iron; D010957:Plasmids; D013302:Streptomyces / Q000235:genetics*
D002021:Buffers; D004269:DNA, Bacterial; D007501:Iron; D004262:DNA Restriction Enzymes