The effect of the chromosome banding techniques on the histone and nonhistone proteins of isolated chromatin

1982 ◽  
Vol 60 (3) ◽  
pp. 328-337 ◽  
Author(s):  
Gary D. Burkholder ◽  
Laurel L. Duczek
Keyword(s):  
Chromosome Banding ◽  
Polyacrylamide Gel Electrophoresis ◽  
Mitotic Chromosomes ◽  
Specific Chromosome ◽  
Nonhistone Proteins ◽  
Chromosomal Proteins ◽  
Different Types ◽  
Protein Alterations ◽  
Chromosome Regions ◽  
Qualitative Changes

The effect of a variety of chromosome banding techniques on the histone and nonhistone proteins of isolated, fixed, air-dried chromatin has been studied. Chromatin preparations were exposed to G-banding (SSC, urea, NaCl–urea, or trypsin), R-banding (Earle's BSS), and C-banding (NaOH or Ba(OH)2) treatments, and the proteins extracted from, and those remaining in the posttreatment chromatin, were examined by SDS polyacrylamide gel electrophoresis. The results indicate that: (i) each banding treatment produces specific quantitative and qualitative changes in the proteins of chromatin; (ii) the diverse treatments producing the same type of chromosome banding have both common and unique effects on the chromosomal proteins; and (iii) the treatments producing different types of chromosome banding have substantially different effects on the chromosomal proteins. It is not known whether these protein alterations are side effects occurring coincidentally with chromosome banding, or whether they are directly involved in mechanisms of banding. Results (ii) and (iii) suggest the latter possibility is more likely. If the protein alterations occur in specific chromosome regions, they may well influence the appearance of bands on mitotic chromosomes.

Fluram induces species-dependent C and G bands in mammalian chromosomes, revealing heterogeneous distribution of chromosomal proteins

Genome ◽  
1991 ◽  
Vol 34 (5) ◽  
pp. 772-776 ◽  
Author(s):  
T. Cuéllar ◽  
J. Gosálvez ◽  
P. Del Castillo ◽  
J. C. Stockert
Keyword(s):  
Banding Pattern ◽  
Chromosome Banding ◽  
Chromosome Region ◽  
Centromeric Heterochromatin ◽  
Primary Amines ◽  
Metaphase Chromosomes ◽  
Heterogeneous Distribution ◽  
Chromosomal Proteins ◽  
Differential Reactivity ◽  
Chromosome Regions

Fluram (Fluorescamine; 4-phenylspiro(furan-2(3H),1′-phthalan)-3,3′-dione) is a fluorogenic reagent, which permits the detection of primary amines by forming highly fluorescent pyrrolinone derivatives. This reagent has been used on methanol – acetic acid fixed metaphase chromosomes of mouse and man and proved to be very effective in differentiating chromosome regions in both genomes. Mouse centromeric heterochromatin is highly reactive, showing intense fluorescence in all centromeric regions, whereas human chromosomes show no fluorescence in such regions. In addition, a G-like banding pattern is also obtained in both types of chromosomes. The differential reactivity of each chromosome region showed by this method demonstrates a heterogeneous distribution of chromosome proteins, resulting in a chromosome banding pattern, which is in this case species dependent.Key words: cytogenetics, chromosome banding, mammalian chromosomes, fluorescence microscopy, Fluram.

scholarly journals Specific glycoprotein changes during development of the chick neural retina.

1978 ◽  
Vol 79 (1) ◽  
pp. 132-137 ◽  
Author(s):  
G Mintz ◽  
L Glaser
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Neural Retina ◽  
Cell Type ◽  
Retinal Antigens ◽  
Qualitative Changes

After separation of whole proteins of chick neural retina by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS), a number of glycoproteins can be detected by staining the gels with 125I-labeled wheat germ agglutinin (WGA) and other lectins. The glycoprotein patterns show both quantitative and qualitative changes between days 7 and 13 of development. Some of these glycoproteins can be separated by chromatography on columns of insolubilized lectins. These observations suggest that purification of some of these glycoproteins identified by staining with radioactive lectins would yield retinal antigens which may be specific for developmental stage and cell type.

Rapid qualitative changes in mRNA populations in cultured human lymphocytes: comparison of the effects of cycloheximide and concanavalin A

1984 ◽  
Vol 62 (9) ◽  
pp. 859-864 ◽  
Author(s):  
Donald R. Forsdyke
Keyword(s):  
Concanavalin A ◽  
Human Lymphocytes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Lymphocyte Activation ◽  
Relative Mass ◽  
Protein Synthesis Inhibitor ◽  
Synthesis Inhibitor ◽  
Mrna Species ◽  
Reticulocyte Lysate ◽  
Qualitative Changes

To examine the hypothesis that the stimulation of cultured lymphocytes by lectins involves the inactivation of a protein repressor of putative "activation genes," the effects of a protein synthesis inhibitor (cycloheximide) and a lectin (concanavalin A) were compared. Qualitative changes in mRNA populations were assessed by translating RNA prepared from cycloheximide- or lectin-treated cultures in a rabbit reticulocyte lysate. [35S]Methionine-labelled translation products were analysed by two-dimensional polyacrylamide gel electrophoresis. Cycloheximide increased the radioactive labelling of cultured lymphocytes with the RNA precursor [3H]uridine, as previously reported. This was observed during the first 3 h of culture; thereafter, cycloheximide was inhibitory. The period of increased labelling with [3H]uridine coincided with a period of great increase in mRNA corresponding to an acidic protein of a relative mass of approximately 55 000. This mRNA was not detected in RNA prepared from control cultures, but was one of the most abundant mRNA species detected in RNA prepared from cycloheximide-treated cultures. Increases in certain less abundant mRNA species were also noted. However, the mRNAs were not observed in RNA prepared from lectin-treated cultures. If an increase in these mRNAs is important for lymphocyte activation, then the increase must be to an extent not detected by our current methods.

The dynamics of individual whey protein concentrations in cows’ mammary secretions during the colostral and early lactation periods

2018 ◽  
Vol 86 (1) ◽  
pp. 88-93 ◽  
Author(s):  
Raquel F.S. Raimondo ◽  
Juliana S.P. Ferrão ◽  
Samantha I. Miyashiro ◽  
Priscila T. Ferreira ◽  
João Paulo E. Saut ◽  
...  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Whey Proteins ◽  
Total Proteins ◽  
Mature Milk ◽  
Rennet Coagulation ◽  
Sds Page ◽  
Different Types ◽  
Biuret Method ◽  
Jersey Cows ◽  
Β Lactoglobulin

AbstractThe bovine whey consists of more than 200 different types of proteins, of which β-lactoglobulin, α-lactalbumin, serum albumin, immunoglobulins and lactoferrin predominate. However, their concentrations are not stable due to the existence of protein dynamics during a transition from colostrum secretion to mature milk. To evaluate the dynamics of whey proteins of Jersey cows during a colostral phase and first month of lactation and an influence of the number of lactations, 268 milk samples from 135 Jersey cows were selected through a clinical evaluation. Whey was obtained by rennet coagulation of the mammary secretion. The concentration of total proteins was determined by the biuret method and their fractions were identified by 12% dodecyl sulfate-polyacrylamide gel electrophoresis (12% SDS-PAGE). Maximum concentrations of all protein fractions were observed in the first 12 h of lactation, reducing over the course of the study. Modification of the protein predominance was also observed. The transition from colostrum secretion to milk occurred between 24 and 72 h postpartum. There was an influence of the number of lactations on the dynamics of whey proteins, indicating that multiparous cows had better immunological and nutritional quality when compared to primiparous cows.

Identification and characterization of cDNA clones encoding two homologous proteins that are part of the asialoglycoprotein receptor

1987 ◽  
Vol 7 (5) ◽  
pp. 1841-1847
Author(s):  
M McPhaul ◽  
P Berg
Keyword(s):  
Rat Liver ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Asialoglycoprotein Receptor ◽  
Cdna Clones ◽  
Differential Splicing ◽  
Homologous Proteins ◽  
Different Types ◽  
Identification And Characterization

The asialoglycoprotein receptor (ASGP-R) from rat liver contains the following three distinct protein species when it is analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis: RHL1 (42 kilodaltons), RHL2 (49 kilodaltons), and RHL3 (54 kilodaltons). In this paper we describe the isolation of cDNA clones encoding RHL1 and RHL2 from a cDNA library constructed from rat liver mRNA. A comparison of the predicted coding sequence for RHL2 with that for RHL1 showed that these sequences are highly homologous. The library also contained numerous cDNA clones for both RHL1 and RHL2 that were derived from unspliced precursor mRNAs. Differential splicing at the 5' end of the RHL1 transcript was inferred from the finding that two different types of RHL1 cDNA were identified, each having a different 5' terminus.

scholarly journals Immunochemical evidence for an additional H-2 region closely linked to H-2D.

1977 ◽  
Vol 145 (2) ◽  
pp. 438-442 ◽  
Author(s):  
T H Hansen ◽  
S E Cullen ◽  
D H Sachs
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Spleen Cell ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Molecular Size ◽  
Migration Patterns ◽  
Different Types ◽  
D Region ◽  
Chemical Evidence

Anti-H-2 reagents have been tested on solubilized spleen cell preparations in combinations expected to be specific for D region products. Two different types of molecules were detected. One showed the expected reactivity with both antisera to private and antisera to public specificities. However, an additional molecule was detected which reacted only with antisera to public specificities. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis migration patterns indicated that both products have a similar molecular size of approximately 45,000 daltons. The data therefore present chemical evidence for the existence of a third H-2-associated gene product of 45,000 mol wt in addition to the classical H-2K and H-2D antigens.

Quinacrine fluorescence of specific chromosome regions

Chromosoma ◽  
1972 ◽  
Vol 36 (4) ◽  
pp. 375-390 ◽  
Author(s):  
J. R. Ellison ◽  
H. J. Barr
Keyword(s):  
Specific Chromosome ◽  
Quinacrine Fluorescence ◽  
Chromosome Regions

Dosage effects on morphological and quantitative traits in maize aneuploids

Genome ◽  
1996 ◽  
Vol 39 (5) ◽  
pp. 898-908 ◽  
Author(s):  
E. A. Lee ◽  
L. L. Darrah ◽  
E. H. Coe
Keyword(s):  
Zea Mays ◽  
Quantitative Traits ◽  
Zea Mays L ◽  
Genetic Relationships ◽  
Specific Chromosome ◽  
Phenotypic Changes ◽  
Dosage Effects ◽  
Quantitative Characters ◽  
Chromosome Arm ◽  
Chromosome Regions

Dosage effects generated by either loss or gain of a chromosome segment were used to identify chromosome regions associated with morphological and quantitative characters in maize (Zea mays L.). Using B–A translocation stocks introgressed into a B73Ht background, a chromosome arm dosage series in a Mo17Ht × B73Ht F1 hybrid background was created for 18 of the 20 chromosome arms. The dosage series was then evaluated for 12 quantitatively inherited characters to associate specific phenotypic changes in a trait with a specific chromosome arm. Not only did our results show the familiar aneuploid syndrome phenomenon, but differential dosage effects among particular chromosome arms were demonstrated. All the quantitative traits measured and all the chromosome arms examined in this study were responsive to changes in chromosome arm dosage. The possible bases behind those differences and their utility in identifying quantitative trait loci, as well as the genetic relationships among the group of quantitatively inherited characters studied, are considered. Key words : corn, chromosome arm, B–A translocations, dosage analysis.

Fractionation of Chromatin Components

1973 ◽  
Vol 51 (5) ◽  
pp. 709-720 ◽  
Author(s):  
John J. Monahan ◽  
Ross H. Hall
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Deae Cellulose ◽  
Polyacrylamide Gel ◽  
Urea Solution ◽  
Equilibrium Density ◽  
Tissue Culture Cell ◽  
Low Molecular Weight ◽  
Nonhistone Proteins

A general method for isolation and fractionation of chromatin into its four major components, DNA, RNA, histories, and nonhistone proteins, is described. The procedure avoids the use of strongly acidic or alkaline conditions, or the use of ionic detergents or phenol. As few as 14 × 106 cells can be used. The procedure is reasonably rapid and has been used successfully with a number of tissue culture cell lines. The chromatin components are dissociated in a 3 M NaCl – 5 M urea solution containing 2-mercaptoethanol and EDTA. The DNA and high molecular weight RNA are collected by high-speed centrifugation and DNA is separated from the RNA by means of Cs2SO4 equilibrium density centrifugation. The histones, nonhistone proteins, and low molecular weight RNA's are fractionated using DEAE-cellulose column chromatography and polyacrylamide gel electrophoresis. A small amount (< 1%) of protein is present in the DNA and RNA fractions. At least 11 low molecular weight RNA subfractions can be detected by means of polyacrylamide gel electrophoresis.

scholarly journals Distribution of protein sulphydryls and disulphides in fixed mammalian chromosomes, and their relationship to banding

1984 ◽  
Vol 70 (1) ◽  
pp. 177-188
Author(s):  
A.T. Sumner
Keyword(s):  
Cross Linking ◽  
Inhomogeneous Distribution ◽  
Mitotic Chromosomes ◽  
Chromosomal Proteins ◽  
C Banding ◽  
Sulphydryl Groups ◽  
Chromosome Bands

Fixed mammalian mitotic chromosomes, when stained with a variety of sensitive, sulphydryl-specific fluorochromes directly or following reduction of disulphide groups, show uniform fluorescence. Thus sulphydryl and disulphide groups are uniformly distributed along the length of the chromosomes, and do not show patterns related to chromosome bands. Performance of G- or C-banding procedures oxidizes sulphydryls to disulphides, but does not produce an inhomogeneous distribution of these groups. Cross-linking sulphydryl groups has no effect on G-, C- or Q-banding of chromosomes. Thus there appears to be no connection between chromosome bands and the distribution or state of oxidation of sulphur in chromosomal proteins.

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