Isolation of Z-DNA-containing plasmids.
Author(s) -
R. Thomae,
Stephan Beck,
Fritz M. Pohl
Publication year - 1983
Publication title -
proceedings of the national academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.80.18.5550
Subject(s) - plasmid , recombinant dna , escherichia coli , dna , microbiology and biotechnology , plasmid preparation , biology , in vitro recombination , dna supercoil , chemistry , molecular cloning , biochemistry , gene , complementary dna , pbr322 , dna replication
A purified, monoclonal antibody, specific for the left-handed Z-form of poly(dG-dC), was coupled covalently to Sephacryl S-1000 beads. Such an antibody column provides a convenient method to isolate and purify those plasmid DNAs that contain Z-DNA from a large excess of other DNAs, RNA, etc. From a library of Escherichia coli DNA, cloned into the vector plasmid pUC-8, several recombinant plasmids were isolated, which bind to this antibody. Thus, E. coli contains sequences, which in "natural" negatively supercoiled DNA, adopt a left-handed Z-DNA-like conformation.
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