Movatterモバイル変換

[0]ホーム
URL:

Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
Thehttps:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

NIH NLM Logo
Log inShow account info
Access keysNCBI HomepageMyNCBI HomepageMain ContentMain Navigation
pubmed logo
Advanced Clipboard
User Guide

Full text links

Atypon full text link Atypon Free PMC article
Full text links

Actions

.2001 Apr;183(7):2219-25.
doi: 10.1128/JB.183.7.2219-2225.2001.

Determination of Wolbachia genome size by pulsed-field gel electrophoresis

Affiliations

Determination of Wolbachia genome size by pulsed-field gel electrophoresis

L V Sun et al. J Bacteriol.2001 Apr.

Abstract

Genome sizes of six different Wolbachia strains from insect and nematode hosts have been determined by pulsed-field gel electrophoresis of purified DNA both before and after digestion with rare-cutting restriction endonucleases. Enzymes SmaI, ApaI, AscI, and FseI cleaved the studied Wolbachia strains at a small number of sites and were used for the determination of the genome sizes of wMelPop, wMel, and wMelCS (each 1.36 Mb), wRi (1.66 Mb), wBma (1.1 Mb), and wDim (0.95 Mb). The Wolbachia genomes studied were all much smaller than the genomes of free-living bacteria such as Escherichia coli (4.7 Mb), as is typical for obligate intracellular bacteria. There was considerable genome size variability among Wolbachia strains, especially between the more parasitic A group Wolbachia infections of insects and the mutualistic C and D group infections of nematodes. The studies described here found no evidence for extrachromosomal plasmid DNA in any of the strains examined. They also indicated that the Wolbachia genome is circular.

PubMed Disclaimer

Figures

FIG. 1
FIG. 1
Composite ethidium bromide-stained gel and corresponding autoradiograph of Southern blot probed with awRiwsp gene fragment. (a) Lane 1, yeast chromosomal size marker; lane 2, undigestedwMelPop genome fragment; lane 3, Southern blot. (b) Lane 1, yeast chromosomal size marker; lane 2, undigestedwMelCS genome fragment; lane 3, undigestedwMel genome fragment; lane 4, Southern blot (undigestedwMelCS genome fragment); lane 5, Southern blot (undigestedwMel genome fragment). (c) Lane 1, yeast chromosomal size marker; lane 2, undigestedwRi genome fragment; lane 3, Southern blot.
FIG. 2
FIG. 2
Autoradiograph of Southern blot probed with mitochondrial 12S rRNA gene fragment. Shown is a DNase I (40 μg/ml) reaction time course at RT (25°C). Lane 1, 15 min; lane 2, 22 min; lane 3, 28 min; lane 4, 35 min; lane 5, 40 min. Arrow,Drosophila mitochondrial genome fragment.
FIG. 3
FIG. 3
Pulsed-field gel sizing ofWolbachia genomes fromD. immitis (a) andB. malayi (b). Lanes 1, ethidium bromide staining of the uncut genomes; lanes 2, corresponding autoradiographs, which were probed with theftsZ gene fragment (a) and with theWolbachia probe cocktail (b); lanes 3, autoradiographs of the genomes after digestion withApaI and probing with theWolbachia probe cocktail. Sizes of selected DNA standards (yeast chromosome and MidRange II pulsed-field gel markers; New England Biolabs) are indicated.
FIG. 4
FIG. 4
CHEF gels of digested genomes of arthropodWolbachia strainswMelPop (a),wMelCS andwMel (b), andwRi (c and d). (a) Lane 1,Saccharomyces cerevisiae chromosomal size marker; lane 2, lambda ladder; lanes 3 to 6, digestedwMelPop genome. (b) Lane 1,S. cerevisiae chromosomal size marker; lane 2, lambda ladder; lane 3, digestedwMelCS genome; lane 4, digestedwMel genome. (c) Lane 1, lambda ladder; lane 2,S. cerevisiae chromosomal size marker; lanes 3 to 6, digestedwRi genome. (d) Lane 1,S. cerevisiae chromosomal size marker; lane 2, lambda ladder; lanes 3 and 4, digestedwRi. Each lane is labeled with the enzyme(s) used.
FIG. 5
FIG. 5
Autoradiographs of Southern blot ofwAlbB probed with awRiwsp gene fragment. Lanes 1 to 3, plugs of Aa23 cells (wAlbB) digested with restriction enzymesAscI (lane 1),FseI (lane 2), andAscI andFseI (lane 3); lane 4, uncut DNA. Arrow,Wolbachia genome fragment that migrated into the gel afterAscI digestion.
See this image and copyright information in PMC

References

    1. Andersson S G, Kurland C G. Reductive evolution of resident genomes. Trends Microbiol. 1998;6:263–268. - PubMed
    1. Ausubel F M, Brent R, Kingston R E, Moore D D, Seidman J G, Smith J A, Struhl K, editors. Current protocols in molecular biology. New York, N.Y: John Wiley & Sons, Inc.; 1994.
    1. Bandi C, Anderson T J, Genchi C, Blaxter M L. Phylogeny of Wolbachia in filarial nematodes. Proc R Soc Lond B Biol Sci. 1998;265:2407–2413. - PMC - PubMed
    1. Bandi C, McCall J W, Genchi C, Corona S, Venco L, Sacchi L. Effects of tetracycline on the filarial worms Brugia pahangi and Dirofilaria immitis and their bacterial endosymbionts Wolbachia. Int J Parasitol. 1999;29:357–364. - PubMed
    1. Baril C, Richaud C, Baranton G, Saint Girons I S, Ferdows M S, Barbour A G. Linear chromosome of Borrelia burgdorferi. Res Microbiol. 1989;140:507–516. - PubMed

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full text links
Atypon full text link Atypon Free PMC article
Cite
Send To

NCBI Literature Resources

MeSHPMCBookshelfDisclaimer

The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). Unauthorized use of these marks is strictly prohibited.

[8]ページ先頭
©2009-2025 Movatter.jp