Characterization of Cloned Mitochondrial DNA from the Teleost Fundulus heteroclitus and its Usefulness as an Interspecies Hybridization Probe

1986 ◽  
Vol 43 (10) ◽  
pp. 1866-1872 ◽  
Author(s):  
Lucia Irene González-Villaseñor ◽  
Amanda M. Burkhoff ◽  
Víctor Corces ◽  
Dennis A. Powers
Keyword(s):  
Mitochondrial Dna ◽  
Ethidium Bromide ◽  
Fundulus Heteroclitus ◽  
Staining Technique ◽  
Hybridization Probe ◽  
Cross Hybridization ◽  
Restriction Patterns ◽  
Ethidium Bromide Staining ◽  
Dna Restriction Fragments ◽  
Dna Restriction

Analysis of mitochondrial DNA endonuclease restriction patterns is a powerful tool for studying related species and variation within species. The ethidium bromide staining technique has limited the number of digestions of mitochondrial DNA per individual. Because 32P-end-labeling also imposes severe limitations, we have resorted to cloning the fish (Fundulus heteroclitus) mitochondrial genome in the lambda replacement vector EMBL-3. The clone was used as a radioactive probe via Southern blotting to detect mitochondrial DNA restriction fragments obtained by multiple restriction endonuclease digestions from small amounts of tissue. This technique offers much greater sensitivity than ethidium bromide staining. Moreover, it eliminates the expense and time to obtain highly purified mitochondrial DNA for the 32P-end-labeling procedure. It is also useful when the mtDNA is prepared from frozen tissue which has been a problem with the 32P-end-labeling technique. Because the cloned mitochondrial DNA has a high degree of cross-hybridization with the mitochondrial DNA of certain other fishes, it can be used to probe the mitochondrial DNA restriction patterns of a variety of fish species. However, its usefulness is restricted by the degree of relatedness to the species being cloned.

Pulsed-field gel electrophoresis applied for comparing Listeria monocytogenes strains involved in outbreaks

1993 ◽  
Vol 39 (4) ◽  
pp. 395-401 ◽  
Author(s):  
C. Buchrieser ◽  
R. Brosch ◽  
B. Catimel ◽  
J. Rocourt
Keyword(s):  
United States ◽  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
The United States ◽  
Pulsed Field Gel Electrophoresis ◽  
Chromosomal Dna ◽  
Restriction Patterns ◽  
Pulsed Field ◽  
Dna Restriction Fragments ◽  
Dna Restriction

Recent food-borne outbreaks of human listeriosis as well as numerous sporadic cases have been mainly caused by Listeria monocytogenes serovar 4b strains. Thus, it was of interest to find out whether a certain clone or a certain few clones were responsible for these cases and especially for outbreaks., We used pulsed-field gel electrophoresis of large chromosomal DNA restriction fragments generated by ApaI, SmaI, or NotI to analyse 75 L. monocytogenes strains isolated during six major and eight smaller recent listeriosis outbreaks. These strains could be divided into 20 different genomic varieties. Thirteen of 14 strains isolated during major epidemics in Switzerland (1983–1987), the United States (California, 1985) and Denmark (1985–1987) demonstrated indistinguishable DNA restriction patterns. In contrast, strains responsible for the outbreaks in Canada (Nova Scotia, 1981), the United States (Massachusetts, 1983), France (Anjou, 1975–1976), New Zealand (1969), and Austria (1986) and some smaller outbreaks in France (1987, 1988, 1989) were each characterized by particular combinations of DNA restriction patterns. Seventy-seven percent of the tested strains could be classified into the previously described ApaI group A (Brosch et al. 1991), demonstrating a very close genomic relatedness. Because 49% of the epidemic strains selected for this study belonged to phagovar 2389/2425/3274/2671/47/108/340 or 2389/47/108/340, fifty-six additional strains of these phagovars, isolated from various origins, were also typed to determine whether differences in DNA restriction profiles between epidemic and randomly selected strains of the same phagovars could be pointed out. Variations in DNA patterns appeared more frequently within randomly selected strains than within epidemic strains.Key words: Listeria monocytogenes, listeriosis, typing, pulsed-field gel electrophoresis, epidemic.

Identification des souches de levures isolées de vins par l'analyse de leur ADN mitochondrial

OENO One ◽  
1987 ◽  
Vol 21 (4) ◽  
pp. 267 ◽  
Author(s):  
Denis Dubourdieu ◽  
Aline Sokol ◽  
Joseph Zucca ◽  
Patrick Thalouarn ◽  
Agnès Dattee ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mitochondrial Dna ◽  
Restriction Patterns ◽  
Adn Mitochondrial ◽  
Practical Applications ◽  
Mise En Œuvre ◽  
Dna Restriction ◽  
Wild Yeasts ◽  
Different Strains

<p style="text-align: justify;">L'analyse des profils de restriction de l'ADN mitochondrial des levures permet une caractérisation fine des souches de <em>Saccharomyces cerevisiae</em>. Cette analyse a été appliquée à deux souches de levures sèches actives et à une vingtaine de souches indigènes, isolées de différents moûts lors de la fermentation spontanée.</p><p style="text-align: justify;">Les profils de restriction de l'ADN mt des souches étudiées présentent une grande diversité.</p><p style="text-align: justify;">La méthode mise en oeuvre est décrite de façon détaillée et les applications pratiques discutées.</p><p style="text-align: justify;">+++</p><p style="text-align: justify;">The analysis of restriction patterns from yeast's mitochondrial DNA leads to a fine characterization of different strains of <em>Saccharomyces cerevisiae</em>. This analysis has been applied to two commercial strains and to twenty wild yeasts, isolated from different musts in case of natural fermentations.</p><p style="text-align: justify;">The DNA restriction patterns of the strains studied present a wide diversity.</p><p style="text-align: justify;">The method we used is minutely described and the practical applications are discussed.</p>

Relatedness of three species of Agaricus inferred from restriction fragment length polymorphism analysis of the ribosomal DNA repeat and mitochondrial DNA

Genome ◽  
1989 ◽  
Vol 32 (2) ◽  
pp. 173-178 ◽  
Author(s):  
William E. A. Hintz ◽  
James B. Anderson ◽  
Paul A. Horgen
Keyword(s):  
Mitochondrial Dna ◽  
Restriction Fragment Length Polymorphism ◽  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
The Other ◽  
Rrna Genes ◽  
Polymorphism Analysis ◽  
Restriction Patterns ◽  
Dna Restriction ◽  
Restriction Fragment Length

The ribosomal DNA (rDNA) repeat of Agaricus brunnescens (= A. bisporus) was cloned and mapped for six restriction endonucleases. The map positions of the 26S, 18S, and 5.8S rRNA genes on the 9.2 kilo base pairs (kbp) repeat were determined by alignment of sites conserved in the rRNA genes of other fungi. The rDNA restriction site maps for six isolates of A. brunnescens, five isolates of A. bitorquis, and three isolates of A. campestris were compared using cloned A. brunnescens (Ag 50) rDNA as a hybridization probe. The rDNA restriction patterns for all six A. brunnescens isolates were identical. The A. bitorquis and A. campestris isolates were subdivided into two groups each, according to rDNA restriction-site polymorphisms. The A. brunnescens and A. bitorquis rDNAs were distinguished by a 0.7 kbp length difference in the noncoding spacer between the 18S and 26S rRNA genes. Despite the almost perfect conservation of the coding region between species, the noncoding spacers of A. campestris and the other two Agaricus species were too divergent to propose a simple series of mutational events to account for the differences. Interstrain and interspecies variation in the mitochondrial DNA was also surveyed. Strain-specific mitochondrial DNA restriction patterns were recognized and fewer differences were observed between the A. brunnescens and A. bitorquis isolates than between A. campestris and the other two species.Key words: Agaricus brunnescens (= A. bisporus), Agaricus, rDNA, mitochondrial DNA, restriction fragment length polymorphism analysis.

CD of ethidium bromide complexes with normal and electrophoretically anomalous DNA restriction fragments

Biopolymers ◽  
1988 ◽  
Vol 27 (7) ◽  
pp. 1107-1126 ◽  
Author(s):  
Elizabeth Slobodyansky ◽  
John Stellwagen ◽  
Nancy C. Stellwagen
Keyword(s):  
Ethidium Bromide ◽  
Restriction Fragments ◽  
Dna Restriction Fragments ◽  
Dna Restriction

Hyperactive recombination in the mitochondrial DNA of the natural death nuclear mutant of Neurospora crassa

1993 ◽  
Vol 13 (11) ◽  
pp. 6778-6788
Author(s):  
H Bertrand ◽  
Q Wu ◽  
B L Seidel-Rogol
Keyword(s):  
Mitochondrial Dna ◽  
Neurospora Crassa ◽  
Nuclear Gene ◽  
General System ◽  
Natural Death ◽  
Dna Restriction Fragments ◽  
Dna Restriction ◽  
Recessive Mutant ◽  
Nuclear Mutant ◽  
Mitochondrial Chromosome

In Neurospora crassa, a recessive mutant allele of a nuclear gene, nd (natural death), causes rapid degeneration of the mitochondrial DNA, a process that is manifested phenotypically as an accelerated form of senescence in growing and stationary mycelia. To examine the mechanisms that are involved in the degradation of the mitochondrial chromosome, several mitochondrial DNA restriction fragments unique to the natural-death mutant were cloned and characterized through restriction, hybridization, and nucleotide sequence analyses. All of the cloned DNA pieces contained one to four rearrangements that were generated by unequal crossing-over between direct repeats of several different nucleotide sequences that occur in pairs and are dispersed throughout the mitochondrial chromosome of wild-type Neurospora strains. The most abundant repeats, a family of GC-rich sequences that includes the so-called PstI palindromes, were not involved in the generation of deletions in the nd mutant. The implication of these results is that the nd allele hyperactivates a general system for homologous recombination in the mitochondria of N. crassa. Therefore, the nd+ allele either codes for a component of the complex of proteins that catalyzes recombination, and possibly repair and replication, of the mitochondrial chromosome or specifies a regulatory factor that controls the synthesis or activity of at least one enzyme or ancillary factor that is affiliated with mitochondrial DNA metabolism.

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