Abstract:
:A technique is described for isolation of plasmid DNA in closed and open circular double-stranded forms from bacterial cells, by use of ATP-dependent deoxyribonuclease purified from Micrococcus luteus. This DNase, acting only upon linear DNA molecules, degrades all bacterial chromosomal DNA extracted in the linear form. Circular plasmid DNAs are left intact, and are then separated by sedimentation through a sucrose gradient. Unlike previous techniques for analysis of plasmid DNA, this technique can be used to isolate not only closed circular DNA but also open circular DNA. Several plasmids, such as those from phage (lambdadv1 and lambdadv21), a colicinogenic factor (Col E2), a sex factor (F(8)' gal), and "minicircles" in Escherichia coli 15, in both the open and closed circular forms, were well separated from chromosomal DNA by this technique.
journal_name
Proc Natl Acad Sci U S Aauthors
Mukai T,Matsubara K,Takagi Ydoi
10.1073/pnas.70.10.2884subject
Has Abstractpub_date
1973-10-01 00:00:00pages
2884-7issue
10eissn
0027-8424issn
1091-6490journal_volume
70pub_type
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