scholarly journals Electrophoretic analysis of Histoplasma capsulatum chromosomal DNA.

1989 ◽  
Vol 9 (3) ◽  
pp. 983-987 ◽  
Author(s):  
P E Steele ◽  
G F Carle ◽  
G S Kobayashi ◽  
G Medoff

Seven chromosome-sized DNA molecules in the Downs strain of Histoplasma capsulatum were resolved by using chromosome-specific DNA probes in blot hybridizations of contour-clamped homogeneous electric field (CHEF) and field-inversion gel electrophoresis (FIGE) agarose gels. The sizes of the chromosomal DNA bands extended from that of the largest Saccharomyces cerevisiae chromosome to beyond that of the Schizosaccharomyces pombe chromosomes. Under our experimental conditions, the order of the five largest DNA bands was inverted in the FIGE gel relative to the CHEF gel, demonstrating a characteristic of FIGE whereby large DNA molecules may have greater rather than lesser mobility with increasing size. Comparison of the Downs strain with other H. capsulatum strains by CHEF and FIGE analysis revealed considerable variability in band mobility. The resolution of seven chromosome-sized DNA molecules in the Downs strain provides a minimum estimate of the chromosome number.

1989 ◽  
Vol 9 (3) ◽  
pp. 983-987
Author(s):  
P E Steele ◽  
G F Carle ◽  
G S Kobayashi ◽  
G Medoff

Seven chromosome-sized DNA molecules in the Downs strain of Histoplasma capsulatum were resolved by using chromosome-specific DNA probes in blot hybridizations of contour-clamped homogeneous electric field (CHEF) and field-inversion gel electrophoresis (FIGE) agarose gels. The sizes of the chromosomal DNA bands extended from that of the largest Saccharomyces cerevisiae chromosome to beyond that of the Schizosaccharomyces pombe chromosomes. Under our experimental conditions, the order of the five largest DNA bands was inverted in the FIGE gel relative to the CHEF gel, demonstrating a characteristic of FIGE whereby large DNA molecules may have greater rather than lesser mobility with increasing size. Comparison of the Downs strain with other H. capsulatum strains by CHEF and FIGE analysis revealed considerable variability in band mobility. The resolution of seven chromosome-sized DNA molecules in the Downs strain provides a minimum estimate of the chromosome number.


1988 ◽  
Vol 8 (4) ◽  
pp. 1469-1473 ◽  
Author(s):  
M J Orbach ◽  
D Vollrath ◽  
R W Davis ◽  
C Yanofsky

A molecular karyotype of Neurospora crassa was obtained by using an alternating-field gel electrophoresis system which employs contour-clamped homogeneous electric fields. The migration of all seven N. crassa chromosomal DNAs was defined, and five of the seven molecules were separated from one another. The estimated sizes of these molecules, based on their migration relative to Schizosaccharomyces pombe chromosomal DNA molecules, are 4 to 12.6 megabases. The seven linkage groups were correlated with specific chromosomal DNA bands by hybridizing transfers of contour-clamped homogeneous electric field gels with radioactive probes specific to each linkage group. The mobilities of minichromosomal DNAs generated from translocation strains were also examined. The methods used for preparation of chromosomal DNA molecules and the conditions for their separation should be applicable to other filamentous fungi.


1988 ◽  
Vol 8 (4) ◽  
pp. 1469-1473 ◽  
Author(s):  
M J Orbach ◽  
D Vollrath ◽  
R W Davis ◽  
C Yanofsky

A molecular karyotype of Neurospora crassa was obtained by using an alternating-field gel electrophoresis system which employs contour-clamped homogeneous electric fields. The migration of all seven N. crassa chromosomal DNAs was defined, and five of the seven molecules were separated from one another. The estimated sizes of these molecules, based on their migration relative to Schizosaccharomyces pombe chromosomal DNA molecules, are 4 to 12.6 megabases. The seven linkage groups were correlated with specific chromosomal DNA bands by hybridizing transfers of contour-clamped homogeneous electric field gels with radioactive probes specific to each linkage group. The mobilities of minichromosomal DNAs generated from translocation strains were also examined. The methods used for preparation of chromosomal DNA molecules and the conditions for their separation should be applicable to other filamentous fungi.


1998 ◽  
Vol 21 (1) ◽  
pp. 11-14 ◽  
Author(s):  
Maria Cléria Valadares-Inglis ◽  
John F. Peberdy

Pulsed-field gel electrophoresis (PFGE) was used to separate chromosome-sized DNA molecules of four strains of Metarhizium anisopliae from Brazil. Metarhizium anisopliae isolates from Japan have been reported as possessing seven chromosomes. Variation was observed among the Brazilian strains and the chromosomal DNA was resolved into eight bands for strain CG46. Densitometric analysis of PFGE gels suggested that the other three Brazilian strains also possess eight chromosomes, with two chromosomes migrating as doublets under the electrophoretic conditions used. The genome size was estimated as varying between 23.39 to 31.88 Mb, not including possible doublet chromosomes.


1994 ◽  
Vol 15 (11) ◽  
pp. 691-696 ◽  
Author(s):  
Carolyn VanCouwenberghe ◽  
Stuart Cohen

AbstractBackground:Xanthomonas maltophiliais increasingly a cause of nosocomial infections. The mode of transmission of this organism is not well known.Objective:To investigate clonality ofX maltophiliaisolates in epidemic and endemic settings.Methods:An outbreak ofX maltophiliawas noted in the Intensive Care Nursery (ICN). Over the ensuing 9 months, hospitalwide isolates ofX maltophiliawere analyzed using contour-clamped homogeneous electric field (CHEF) gel electrophoresis of chromosomal DNA. This method was compared with the antibiogram for detecting differences and similarities among strains.Results:X maltophiliawas recovered from 76 sites in 72 patients; 65 isolates from 61 patients and the hands of one nurse were available for analysis. CHEF demonstrated differences between most epidemiologically unrelated strains and similarity between most epidemiologically related strains. Several strains, initially presumed to be related because of temporal and spatial proximity of the patients involved, were determined by CHEF analysis to be independent infections. One pair of isolates whoseXbalCHEF patterns differed by a single band were differentiated clearly by Sspl. There was enough variation in the minimum inhibitory concentrations of selected antibiotics to allow typing of some strains. The antibiogram, however, did not group all of the ICN outbreak isolates with others found to be genetically identical by CHEF and it grouped 39 of 56 isolates with others not genetically the same.Conclusions:Although it is a convenient and economical tool, the antibiogram has limitations. Analysis by CHEF should help to elucidate the epidemiological spread ofX maltophiliain the hospital.


2020 ◽  
Vol 14 (4) ◽  
pp. 044114
Author(s):  
Ryo Inukai ◽  
Hidekuni Takao ◽  
Fusao Shimokawa ◽  
Kyohei Terao

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