scholarly journals Macronuclear DNA molecules of Tetrahymena thermophila.

1986 ◽  
Vol 6 (3) ◽  
pp. 900-905 ◽  
Author(s):  
R K Conover ◽  
C F Brunk
Keyword(s):  
Gel Electrophoresis ◽  
Molecular Mechanisms ◽  
Tetrahymena Thermophila ◽  
Fragmentation Process ◽  
Dna Molecules ◽  
Physical Organization ◽  
Different Strains ◽  
Average Size ◽  
Orthogonal Field

The physical organization of the DNA in the macronuclei of Tetrahymena thermophila was investigated by using alternating-orthogonal-field gel electrophoresis. The genome consisted of a spectrum of molecules with lengths ranging from less than 100 to in excess of 1,500 kilobase pairs. There were about 270 different macronuclear DNA molecules, with an average size of about 800 kilobase pairs. Specific genes were mapped and were generally found on macronuclear DNA molecules of the same size in different strains of T. thermophila. This indicates that the molecular mechanisms giving rise to the macronuclear DNA molecules were precise. The fragmentation process that gave rise to macronuclear DNA molecules occurred between 11 and 19 h after the initiation of conjugation.

Macronuclear DNA molecules of Tetrahymena thermophila

1986 ◽  
Vol 6 (3) ◽  
pp. 900-905
Author(s):  
R K Conover ◽  
C F Brunk
Keyword(s):  
Gel Electrophoresis ◽  
Molecular Mechanisms ◽  
Tetrahymena Thermophila ◽  
Fragmentation Process ◽  
Dna Molecules ◽  
Physical Organization ◽  
Different Strains ◽  
Average Size ◽  
Orthogonal Field

The physical organization of the DNA in the macronuclei of Tetrahymena thermophila was investigated by using alternating-orthogonal-field gel electrophoresis. The genome consisted of a spectrum of molecules with lengths ranging from less than 100 to in excess of 1,500 kilobase pairs. There were about 270 different macronuclear DNA molecules, with an average size of about 800 kilobase pairs. Specific genes were mapped and were generally found on macronuclear DNA molecules of the same size in different strains of T. thermophila. This indicates that the molecular mechanisms giving rise to the macronuclear DNA molecules were precise. The fragmentation process that gave rise to macronuclear DNA molecules occurred between 11 and 19 h after the initiation of conjugation.

Imaging of kinked configurations of DNA molecules undergoing orthogonal field alternating gel electrophoresis by fluorescence microscopy

Biochemistry ◽  
1990 ◽  
Vol 29 (13) ◽  
pp. 3396-3401 ◽  
Author(s):  
Sergio Gurrieri ◽  
Enrico Rizzarelli ◽  
David Beach ◽  
Carlos Bustamante
Keyword(s):  
Fluorescence Microscopy ◽  
Gel Electrophoresis ◽  
Dna Molecules ◽  
Orthogonal Field

scholarly journals Purification and staining of intact yeast DNA chromosomes and real-time observation of their migration during gel electrophoresis

1997 ◽  
Vol 326 (1) ◽  
pp. 131-138 ◽  
Author(s):  
Sergio GURRIERI ◽  
Carlos BUSTAMANTE
Keyword(s):  
Gel Electrophoresis ◽  
Molecular Mechanisms ◽  
Pulsed Field Gel Electrophoresis ◽  
Dna Molecules ◽  
Shear Forces ◽  
The Past ◽  
Intermediate Size ◽  
Time Observation ◽  
Dna Trapping ◽  
Real Time Observation

In the past few years, fluorescence microscopy has been used successfully to characterize the motion of intermediate-size DNA molecules (50–500 kbp) during steady- and pulsed-field gel electrophoresis. However, experimental difficulties had prevented the application of this technique to the direct observation of longer DNA chromosomes (1–2 Mbp). In the present study a particular procedure was followed for the purification and staining of chromosomal yeast DNA to protect it from shear forces. Also, a new highly fluorescent DNA-labelling dye, YOYO-1, was employed to improve brightness and contrast. Finally, the motion of such long DNA molecules (1–2 Mbp) was characterized under steady-field electrophoresis conditions. An accurate description of the molecular mechanisms of motion of such long molecules should provide the basis for a detailed analysis of the mechanisms responsible for DNA trapping.

Assignment of cloned genes to the seven electrophoretically separated Candida albicans chromosomes

1988 ◽  
Vol 8 (11) ◽  
pp. 4721-4726
Author(s):  
B B Magee ◽  
Y Koltin ◽  
J A Gorman ◽  
P T Magee
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Candida Albicans ◽  
Unknown Function ◽  
Gel Electrophoresis ◽  
Homogeneous Field ◽  
Electrophoretic Karyotype ◽  
Chromosome Size ◽  
Homologous Chromosomes ◽  
Orthogonal Field ◽  
Chromosomal Bands

By using orthogonal-field alternating gel electrophoresis (OFAGE), field-inversion gel electrophoresis (FIGE), and contour-clamped homogeneous field gel electrophoresis (CHEF), we have clearly resolved 11 chromosomal bands from various Candida albicans strains. OFAGE resolves the smaller chromosomes better, while FIGE, which under our conditions causes the chromosomes to run in the reverse order of OFAGE, is more effective in separating the larger chromosomes. CHEF separates all chromosomes under some conditions, but these conditions do not often resolve homologs. The strains examined are highly polymorphic for chromosome size. Fourteen cloned Candida genes, isolated on the basis of conferral of new properties to or complementation of auxotrophic deficiencies in Saccharomyces cerevisiae, and three sequences of unknown function have been hybridized to Southern transfers of CHEF, FIGE, and OFAGE gels. Four sets of resolvable bands have been shown to be homologous chromosomes. On the basis of these data, we suggest that C. albicans has seven chromosomes. Genes have been assigned to the seven chromosomes. Two chromosomes identified genetically have been located on the electrophoretic karyotype.

Separation of Large DNA Molecules, Greater than 25 Kbp, by Gel Electrophoresis

1989 ◽  
pp. 71-84
Author(s):  
John F. T. Spencer ◽  
Dorothy M. Spencer ◽  
I. J. Bruce
Keyword(s):  
Gel Electrophoresis ◽  
Dna Molecules
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