Direct end labelling of telomeres

Genome ◽  
1993 ◽  
Vol 36 (2) ◽  
pp. 224-229 ◽  
Author(s):  
Alexander Kolchinsky ◽  
Peter M. Gresshoff
Keyword(s):  
Glycine Max ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Telomeric Sequence ◽  
Two Dimensional ◽  
Pulsed Field ◽  
Two Dimensional Electrophoresis ◽  
Restriction Nuclease ◽  
Novel Approach ◽  
Dimensional Electrophoresis

A novel approach of direct end labelling of telomeres is presented. Chromosome-sized, agarose-embedded DNA was treated with T4 DNA polymerase to remove protruding 3′ end of telomeres and to generate single-stranded 5′ ends. The DNA was then labelled by the same enzyme in the presence of [α-32P|dGTP and cold dATP and dTTP. Labelled yeast chromosomes separated by pulsed field gel electrophoresis maintained their integrity. Digestion of yeast chromosomes separated in pulsed field gels with a restriction nuclease (HinfI), followed by conventional electrophoresis in the second dimension, resulted in a fingerprint-like pattern of labelled telomeres. This was very similar to the hybridization pattern of a similar two-dimensional gel probed with cloned yeast telomeric sequence. The same approach enabled us to label telomeres in soybean, determine their size, and to reveal polymorphisms in the length of telomeres between the closely related subspecies Glycine max (soybean) and Glycine soja.Key words: telomeres, Saccharomyces cerevisiae, Glycine max, two-dimensional electrophoresis, DNA polymorphism.

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.

Assessment of the protein quality of fourteen soybean [Glycine max (L.) Merr.] cultivars using amino acid analysis and two-dimensional electrophoresis

2007 ◽  
Vol 40 (1) ◽  
pp. 129-146 ◽  
Author(s):  
Constantinos G. Zarkadas ◽  
Christine Gagnon ◽  
Stephen Gleddie ◽  
Shahrokh Khanizadeh ◽  
Elroy R. Cober ◽  
...  
Keyword(s):  
Glycine Max ◽  
Amino Acid ◽  
Amino Acid Analysis ◽  
Protein Quality ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Glycine Max L ◽  
Dimensional Electrophoresis

Evaluation of lipoproteins and apolipoproteins in serum of a Tangier patient by micro-scale two-dimensional electrophoresis.

1987 ◽  
Vol 33 (4) ◽  
pp. 468-472 ◽  
Author(s):  
T Manabe ◽  
S Visvikis ◽  
M F Dumon ◽  
M Clerc ◽  
G Siest
Keyword(s):  
Molecular Mass ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Disease Patient ◽  
High Density Lipoproteins ◽  
Two Dimensional ◽  
High Concentration ◽  
Two Dimensional Electrophoresis ◽  
Micro Scale ◽  
Dimensional Electrophoresis

Abstract We examined lipoproteins and apolipoproteins in serum of a Tangier-disease patient. We used three different techniques of micro-scale two-dimensional electrophoresis: (a) no denaturants; (b) with sodium dodecyl sulfate (SDS) used only in the slab gel electrophoresis; (c) and with urea and a detergent used in isoelectric focusing and with SDS in slab gel electrophoresis. By technique a, an extremely low concentration of high-density lipoproteins (HDL) in the Tangier serum was seen, and lipoproteins that cannot form HDL complexes were detected as multiple spots in the acidic (pl 4 approximately 5) and relatively low apparent molecular mass (20,000 approximately 80,000) region. By technique b, Tangier low-molecular-mass lipoproteins were dissociated into their constituent apolipoproteins, and we observed a higher proportion of apoC-III, together with lower proportions of apoA-I and apoA-II, than in the normal HDL fraction. Technique c showed the total content of apolipoproteins in the whole Tangier serum, as several workers have reported. The presence of low-molecular-mass lipoproteins and a high concentration of apoC-III in this lipoprotein fraction characterized the Tangier serum.

THE NATURE OF TWO COMPONENTS OF PIG MUCOSAL HEPARIN, SEPARATED BY ELECTROPHORESIS IN AGAROSE GEL

1967 ◽  
Vol 45 (7) ◽  
pp. 1015-1020 ◽  
Author(s):  
Alan A. Horner
Keyword(s):  
Gel Electrophoresis ◽  
Nitrous Acid ◽  
Anticoagulant Activity ◽  
Thin Layers ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
Uronic Acids ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis

Two components of pig mucosal heparin were separated by agarose-gel electrophoresis and recovered from the gels. The slower moving component had the higher anticoagulant activity and intrinsic viscosity. The faster moving component had the higher sulfate : carboxyl ratio. Each component was degraded by self-hydrolysis to N-desulfated heparin, deaminated with nitrous acid, and hydrolyzed with formic acid. Two-dimensional electrophoresis and chromatography on thin layers of cellulose showed that each component had two uronic acids with the characteristics of glucuronic and iduronic acids.

Two-dimensional motion of DNA bands during 120° pulsed-field gel electrophoresis. I. Effect of molecular weight

Biopolymers ◽  
1995 ◽  
Vol 35 (3) ◽  
pp. 297-306 ◽  
Author(s):  
M. Shane Hutson ◽  
George Holzwarth ◽  
Thomas Duke ◽  
Jean-Louis Viovy
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Two Dimensional ◽  
Pulsed Field
Sign in / Sign up

Export Citation Format

Share Document