RRC ID |
3233
|
著者 |
Adachi S, Kohiyama M, Onogi T, Hiraga S.
|
タイトル |
Localization of replication forks in wild-type and mukB mutant cells of Escherichia coli.
|
ジャーナル |
Mol Genet Genomics
|
Abstract |
To examine the subcellular localization of the replication machinery in Escherichia coli, we have developed an immunofluorescence method that allows us to determine the subcellular location of newly synthesized DNA pulse-labeled with 5-bromo-2'-deoxyuridine (BrdU). Using this technique, we have analyzed growing cells. In wild-type cells that showed a single BrdU fluorescence signal, the focus was located in the middle of the cell; in cells with two signals, the foci were localized at positions equivalent to 1/4 and 3/4 of the cell length. The formation of BrdU foci was dependent upon ongoing chromosomal replication. A mutant lacking MukB, which is required for proper partitioning of sister chromosomes, failed to maintain the ordered localization of BrdU foci: (1) a single BrdU focus tended to be localized at a pole-proximal region of the nucleoid, and (2) a focus was often found to consist of two replicating chromosomes. Thus, the positioning of replication forks is affected by the disruption of the mukB gene.
|
巻・号 |
274(3)
|
ページ |
264-71
|
公開日 |
2005-10-1
|
DOI |
10.1007/s00438-005-0023-6
|
PMID |
16133165
|
MeSH |
Bromodeoxyuridine
Chromosomal Proteins, Non-Histone / genetics*
DNA / genetics*
DNA / metabolism
DNA Replication / genetics*
Escherichia coli / genetics*
Escherichia coli Proteins / genetics*
Flow Cytometry
Fluorescent Antibody Technique, Indirect / methods
Immunohistochemistry
|
IF |
2.797
|
引用数 |
19
|
WOS 分野
|
GENETICS & HEREDITY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
原核生物(大腸菌) |
YK1100
AZ5372
ME8775(PC2) |