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Use of site-specific recombination as a probe of DNA structure and metabolism in vivo.

Abstract：

:We used site-specific recombination catalyzed by the bacteriophage lambda Int system to probe DNA structure and metabolism in vivo. In vitro, the complexity of catenated products was linearly proportional to substrate supercoil density. A system was developed that gave efficient, controlled Int recombination in Escherichia coli cells. From a comparison of the data obtained in vitro and in vivo, we conclude that Int recombination does have the same mechanism in vivo as it has in vitro, but that only 40% of the plasmid DNA linking deficit in E. coli cells may be in the interwound supercoil form demonstrated in vitro. We suggest that this is the effective level of supercoiling in vivo, because the remaining DNA is constrained in alternative forms by protein binding. The study of Int recombination in vivo also provides an assay for enzymes that decatenate circular molecules, such as those formed during DNA replication. We find that DNA gyrase is the principal decatenase in E. coli and that it acts spontaneously and rapidly.

journal_name

J Mol Biol

journal_title

Journal of molecular biology

authors

Bliska JB,Cozzarelli NR

doi

10.1016/0022-2836(87)90369-x

subject

Has Abstract

pub_date

1987-03-20 00:00:00

pages

205-18

issue

2

eissn

0022-2836

issn

1089-8638

pii

0022-2836(87)90369-X

journal_volume

194

pub_type

杂志文章

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