A physical and genetic map of theMycoplasma hyopneumoniaestrain J genome

2000 ◽  
Vol 46 (9) ◽  
pp. 832-840 ◽  
Author(s):  
Walter A Blank ◽  
Gerald W Stemke
Keyword(s):  
Genetic Map ◽  
Dna Hybridization ◽  
Gel Electrophoresis ◽  
Type Strain ◽  
Physical Map ◽  
Mycoplasma Hyopneumoniae ◽  
Pulsed Field Gel Electrophoresis ◽  
Cosmid Library ◽  
Physical And Genetic Map ◽  
Genomic Map

A macrorestriction map of the genome of Mycoplasma hyopneumoniae strain J, the type strain of the causative agent of enzootic pneumonia in pigs, was constructed using pulsed-field gel electrophoresis (PFGE) and DNA hybridization. The size of the genome as determined by PFGE was approximately 1070 kb. Assembly of the M. hyopneumoniae genomic map was facilitated and complimented by the simultaneous construction of an ordered cosmid library. Five contigs of overlapping cosmids were assembled, which together represent coverage of approximately 728 kb. Forty-two genetic markers (including three types of repeated elements) were placed on the M. hyopneumoniae map. Closer examination of an ApaI restriction fragment contained entirely within a single cosmid insert suggests that the genome size may be overestimated by PFGE.Key words: Mycoplasma hyopneumoniae, mollicutes, physical map, genetic map.

Physical and genetic map of the Spiroplasma kunkelii CR2-3x chromosome

2006 ◽  
Vol 52 (9) ◽  
pp. 857-867 ◽  
Author(s):  
Ellen L Dally ◽  
Thereza S.L Barros ◽  
Yan Zhao ◽  
ShaoPing Lin ◽  
Bruce A Roe ◽  
...  
Keyword(s):  
Genetic Map ◽  
Gel Electrophoresis ◽  
Southern Hybridization ◽  
Pulsed Field Gel Electrophoresis ◽  
Enzyme Analysis ◽  
Two Dimensional ◽  
Pulsed Field ◽  
Spiroplasma Kunkelii ◽  
Corn Stunt ◽  
Physical And Genetic Map

Spiroplasma kunkelii (class Mollicutes) is the characteristically helical, wall-less bacterium that causes corn stunt disease. A combination of restriction enzyme analysis, pulsed-field gel electrophoresis (PFGE), and Southern hybridization analysis was used to construct a physical and genetic map of the S. kunkelii CR2-3x chromosome. The order of restriction fragments on the map was determined by analyses of reciprocal endonuclease double digests employing I-CeuI, AscI, ApaI, EagI, SmaI, BssHII, BglI, and SalI; adjacent fragments were identified on two-dimensional pulsed-field electrophoresis gels. The size of the chromosome was estimated at 1550 kb. Oligonucleotide pairs were designed to prime the amplification of 26 S. kunkelii gene sequences in the polymerase chain reaction (PCR). Using PCR amplicons as probes, the locations of 27 S. kunkelii putative single-copy genes were positioned on the map by Southern hybridization analyses of chromosomal fragments separated in PFGE. The nucleotide sequence of the single ribosomal RNA operon was determined and its location mapped to a chromosomal segment bearing recognition sites for SalI, SmaI, EagI, and I-CeuI.Key words: Spiroplasma kunkelii CR2-3x, corn stunt spiroplasma, mollicutes, genome mapping, two-dimensional pulsed-field gel electrophoresis.

scholarly journals Development ofa Physical and Genetic Map of the Virulent WolbachiaStrainwMelPop

2003 ◽  
Vol 185 (24) ◽  
pp. 7077-7084 ◽  
Author(s):  
Ling V. Sun ◽  
Markus Riegler ◽  
Scott L. O'Neill
Keyword(s):  
Genetic Map ◽  
Gel Electrophoresis ◽  
Southern Hybridization ◽  
Gene Organization ◽  
Pulsed Field Gel Electrophoresis ◽  
Whole Genome Sequence ◽  
Whole Genome ◽  
Restriction Fragments ◽  
Chromosome Fragments ◽  
Physical And Genetic Map

ABSTRACT We report here the construction of a physical and genetic map of the virulent Wolbachia strain, wMelPop. This map was determined by ordering 28 chromosome fragments that resulted from digestion with the restriction endonucleases FseI, ApaI, SmaI, and AscI and were resolved by pulsed-field gel electrophoresis. Southern hybridization was done with 53 Wolbachia-specific genes as probes in order to determine the relative positions of these restriction fragments and use them to serve as markers. Comparison of the resulting map with the whole genome sequence of the closely related benign Wolbachia strain, wMel, shows that the two genomes are largely conserved in gene organization with the exception of a single inversion in the chromosome.

Construction of a genomic map ofMoraxella (Branhamella) catarrhalisATCC 25238 and physical mapping of virulence-associated genes

1999 ◽  
Vol 45 (4) ◽  
pp. 299-303 ◽  
Author(s):  
K T Nguyen ◽  
E J Hansen ◽  
M A Farinha
Keyword(s):  
Gel Electrophoresis ◽  
Moraxella Catarrhalis ◽  
Southern Hybridization ◽  
Physical Map ◽  
Outer Membrane Proteins ◽  
Restriction Enzymes ◽  
Pulsed Field Gel Electrophoresis ◽  
Respiratory Pathogens ◽  
Circular Chromosome ◽  
Pulsed Field

A physical genome map of the Moraxella catarrhalis type strain (ATCC 25238) has been constructed using pulsed field gel electrophoresis. Macrorestriction analyses of the genome of M. catarrhalis were performed by digestion with the restriction enzymes SmaI, NotI, and RsrII, which cleave the single circular chromosome into 9, 10, and 6 fragments, respectively. The chromosomal fragments generated by pulsed field gel electrophoresis were converted to a linkage map utilizing a combination of partial digestions, and cross-hybridizations. Moraxella catarrhalis, like a number of other respiratory pathogens, has a relatively small genome estimated at 1750 kilobase pairs or about 40% of the size of the Escherichia coli genome. The locations of the four ribosomal RNA operons (rrnLS) were determined by Southern hybridization and by digestion with I-CeuI endonuclease. A number of genes involved in virulence have been placed onto the physical map by Southern hybridization including those encoding the predominant outer-membrane proteins and the chromosomal gene encoding beta-lactamase.Key words: Moraxella catarrhalis, physical map, genome analysis, pulsed-field gel electrophoresis, virulence.

scholarly journals Use of pulsed field gel electrophoresis and transposon mutagenesis to estimate the minimal number of genes required for motility in Caulobacter crescentus.

Genetics ◽  
1989 ◽  
Vol 123 (4) ◽  
pp. 649-654 ◽  
Author(s):  
B Ely ◽  
T W Ely
Keyword(s):  
Gel Electrophoresis ◽  
Caulobacter Crescentus ◽  
Pulsed Field Gel Electrophoresis ◽  
Restriction Fragments ◽  
Pulsed Field ◽  
Transposon Insertion ◽  
Number Of Genes ◽  
Physical And Genetic Map ◽  
Insertion Mutations ◽  
Flagellar Genes

Abstract To facilitate the mapping of transposon insertion mutations in Caulobacter crescentus, we have used pulsed field gel electrophoresis to construct a detailed physical and genetic map of the C. crescentus genome. Restriction fragments were generated by DraI, AseI, or SpeI which cleave the C. crescentus 40, 13, and 26 times, respectively, and Tn5 insertions were used to align the restriction fragments generated by each of the enzymes. The utility of the resulting map was demonstrated by determining the chromosomal locations of a collection of flagellar mutations. As a result of this study, we were able to identify ten new flagellar genes at various locations on the chromosome. Thus, at least 48 genes are required for the assembly of a functional flagellum in C. crescentus.

scholarly journals A physical map of the human regulator of complement activation gene cluster linking the complement genes CR1, CR2, DAF, and C4BP.

1988 ◽  
Vol 167 (2) ◽  
pp. 664-669 ◽  
Author(s):  
J Rey-Campos ◽  
P Rubinstein ◽  
S Rodriguez de Cordoba
Keyword(s):  
Gene Cluster ◽  
Complement Activation ◽  
Gel Electrophoresis ◽  
Physical Map ◽  
Pulsed Field Gel Electrophoresis ◽  
Complement Components ◽  
Pulsed Field ◽  
Human Genes ◽  
Tight Linkage ◽  
Genes Encoding

We report the organization of the human genes encoding the complement components C4-binding protein (C4BP), C3b/C4b receptor (CR1), decay accelerating factor (DAF), and C3dg receptor (CR2) within the regulator of complement activation (RCA) gene cluster. Using pulsed field gel electrophoresis analysis these genes have been physically linked and aligned as CR1-CR2-DAF-C4BP in an 800-kb DNA segment. The very tight linkage between the CR1 and the C4BP loci, contrasted with the relative long DNA distance between these genes, suggests the existence of mechanisms interfering with recombination within the RCA gene cluster.

scholarly journals Australian Isolates of Legionella longbeachae Are Not a Clonal Population

1999 ◽  
Vol 37 (10) ◽  
pp. 3249-3254 ◽  
Author(s):  
J. C. Montanaro-Punzengruber ◽  
L. Hicks ◽  
W. Meyer ◽  
G. L. Gilbert
Keyword(s):  
Genetic Variability ◽  
Legionella Pneumophila ◽  
Gel Electrophoresis ◽  
Type Strain ◽  
Pulsed Field Gel Electrophoresis ◽  
Epidemiological Investigation ◽  
Environmental Isolates ◽  
Clonal Population ◽  
Pulsed Field ◽  
Major Cluster

Legionella longbeachae is almost as frequent a cause of legionellosis in Australia as Legionella pneumophila, but epidemiological investigation of possible environmental sources and clinical cases has been limited by the lack of a discriminatory subtyping method. The purpose of this study was to examine the genetic variability among Australian isolates of L. longbeachaeserogroup 1. Pulsed-field gel electrophoresis (PFGE) ofSfiI fragments revealed three distinct pulsotypes among 57 clinical and 11 environmental isolates and the ATCC control strains of L. longbeachae serogroups 1 and 2. Each pulsotype differed by four bands, corresponding to <65% similarity. A clonal subgroup within each pulsotype was characterized by >88% similarity. The largest major cluster was pulsotype A, which included 43 clinical isolates and 9 environmental isolates and was divided into five subgroups. Pulsotypes B and C comprised smaller numbers of clinical and environmental isolates, which could each be further divided into three subgroups. The ATCC type strain of L. longbeachae serogroup 1 was classified as pulsotype B, subtype B3, while the ATCC type strain of L. longbeachae serogroup 2 was identified as a different pulsotype, LL2. SfiI macrorestriction analysis followed by PFGE showed that the AustralianL. longbeachae strains are not a single clonal population as previously reported.

scholarly journals Unconventional Genomic Organization in the Alpha Subgroup of the Proteobacteria

1998 ◽  
Vol 180 (10) ◽  
pp. 2749-2755 ◽  
Author(s):  
Estelle Jumas-Bilak ◽  
Sylvie Michaux-Charachon ◽  
Gisele Bourg ◽  
Michel Ramuz ◽  
Annick Allardet-Servent
Keyword(s):  
Dna Hybridization ◽  
Gel Electrophoresis ◽  
Genomic Organization ◽  
Pulsed Field Gel Electrophoresis ◽  
Point Of View ◽  
Gene Probe ◽  
Ochrobactrum Anthropi ◽  
Bartonella Quintana ◽  
The Family ◽  
Taxonomic Point

ABSTRACT Pulsed-field gel electrophoresis was used to analyze the genomic organization of 16 bacteria belonging or related to the familyRhizobiaceae of the alpha subgroup of the classProteobacteria. The number and sizes of replicons were determined by separating nondigested DNA. Hybridization of anrrn gene probe was used to distinguish between chromosomes and plasmids. Members of the genus Agrobacterium all possess two chromosomes, and each biovar has a specific genome size. As previously demonstrated for Agrobacterium tumefaciens C58, the smaller chromosomes of Agrobacterium biovar 1 andAgrobacterium rubi strains appear to be linear. The genomes of Rhizobium strains were all of similar sizes but were seen to contain either one, two, or three megareplicons. Only one chromosome was present in the member of the related genusPhyllobacterium. We found one or two chromosomes inRhodobacter and Brucella species, two chromosomes in Ochrobactrum anthropi, and one chromosome inMycoplana dimorpha and Bartonella quintana; all of these genera are related to the Rhizobiaceae. The presence of multiple chromosomes is discussed from a phylogenetic and taxonomic point of view.

Many yeast chromosomes lack the telomere-specific Y' sequence

1989 ◽  
Vol 9 (12) ◽  
pp. 5754-5757
Author(s):  
D Jäger ◽  
P Philippsen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Hybridization ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Different Strains

Chromosomal DNAs of 26 different strains representing Saccharomyces species were analyzed by pulsed-field gel electrophoresis and subsequent hybridization to Y' telomere DNA. Hybridization to Y' was found exclusively in Saccharomyces cerevisiae strains, and among these strains, Y' sequences were found to be lacking in small, middle-sized, and large chromosomes.

scholarly journals Many yeast chromosomes lack the telomere-specific Y' sequence.

1989 ◽  
Vol 9 (12) ◽  
pp. 5754-5757 ◽  
Author(s):  
D Jäger ◽  
P Philippsen
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Hybridization ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Different Strains

Chromosomal DNAs of 26 different strains representing Saccharomyces species were analyzed by pulsed-field gel electrophoresis and subsequent hybridization to Y' telomere DNA. Hybridization to Y' was found exclusively in Saccharomyces cerevisiae strains, and among these strains, Y' sequences were found to be lacking in small, middle-sized, and large chromosomes.

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