Chromosome preparations from protoplasts: in situ hybridization banding pattern of a dispersed DNA sequence in rye (Secale cereale L.)

Genome ◽  
1991 ◽  
Vol 34 (4) ◽  
pp. 524-527 ◽  
Author(s):  
N. Jouve ◽  
C. L. McIntyre ◽  
J. P. Gustafson
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Banding Pattern ◽  
Banding Technique ◽  
Differential Distribution ◽  
Cellular Debris ◽  
Biotin Labeling ◽  
Secale Cereale L

The utilization of genome-specific DNA sequences coupled with in situ hybridization for chromosome karyotyping in wheat, rye, and triticale has been of limited value because of the presence of cellular and cytoplasmic debris. The use of protoplasts, thus eliminating cellular debris, has been shown to improve the level of detection of low-copy and unique DNA sequences in cereals. Therefore, the use of protoplasts could represent an appropriate tool to improve the results of karyotyping cereal chromosomes with genome-specific DNA sequences. This paper describes the results on the comparative application of protoplasts and squash preparations in the analysis of physical mapping of a dispersed DNA sequence (pSc119.1) to rye chromosomes by in situ hybridization. Individual chromosomes of rye were not distinguishable by their hybridization patterns to pSc119.1 when squash preparations were used. These showed an undefined distribution of the DNA probe that covered apparently the entire length of each rye chromosome. However, considerable improvement was observed for the differential distribution of the pSc119.1 DNA sequence in protoplast preparations. The karyotypic banding pattern of pSc119.1 showed a better banding pattern than can be observed using the C-banding technique. Therefore, the use of protoplasts hybridized with dispersed DNA markers could be of more value in monitoring chromosome karyotypes than existing cytological techniques.Key words: biotin labeling, dispersed sequences, rye.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Identification of the entire chromosome complement of bread wheat by two-colour FISH

Genome ◽  
1997 ◽  
Vol 40 (5) ◽  
pp. 589-593 ◽  
Author(s):  
C. Pedersen ◽  
P. Langridge
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Dna Sequences ◽  
Banding Pattern ◽  
Aegilops Tauschii ◽  
Chromosome Complement ◽  
Chromosome Identification ◽  
Satellite Sequence ◽  
Entire Chromosome

Using the Aegilops tauschii clone pAs1 together with the barley clone pHvG38 for two-colour fluorescence in situ hybridization (FISH) the entire chromosome complement of hexaploid wheat was identified. The combination of the two probes allowed easy discrimination of the three genomes of wheat. The banding pattern obtained with the pHvG38 probe containing the GAA-satellite sequence was identical to the N-banding pattern of wheat. A detailed idiogram was constructed, including 73 GAA bands and 48 pAs1 bands. Identification of the wheat chromosomes by FISH will be particularly useful in connection with the physical mapping of other DNA sequences to chromosomes, or for chromosome identification in general, as an alternative to C-banding.Key words: Triticum aestivum, chromosome identification, fluorescence in situ hybridization, repetitive DNA sequences.

Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research

Genome ◽  
2006 ◽  
Vol 49 (9) ◽  
pp. 1057-1068 ◽  
Author(s):  
Jiming Jiang ◽  
Bikram S. Gill
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Molecular Cytogenetics ◽  
Plant Genome ◽  
Current Status ◽  
Sequence Information ◽  
Resolution Mapping

Fluorescence in situ hybridization (FISH), which allows direct mapping of DNA sequences on chromosomes, has become the most important technique in plant molecular cytogenetics research. Repetitive DNA sequence can generate unique FISH patterns on individual chromosomes for karyotyping and phylogenetic analysis. FISH on meiotic pachytene chromosomes coupled with digital imaging systems has become an efficient method to develop physical maps in plant species. FISH on extended DNA fibers provides a high-resolution mapping approach to analyze large DNA molecules and to characterize large genomic loci. FISH-based physical mapping provides a valuable complementary approach in genome sequencing and map-based cloning research. We expect that FISH will continue to play an important role in relating DNA sequence information to chromosome biology. FISH coupled with immunoassays will be increasingly used to study features of chromatin at the cytological level that control expression and regulation of genes.

Fluorescence in situ hybridization on plant extended chromatin DNA fibers for single-copy and repetitive DNA sequences

2011 ◽  
Vol 30 (9) ◽  
pp. 1779-1786 ◽  
Author(s):  
Kun Yang ◽  
Hecui Zhang ◽  
Richard Converse ◽  
Yong Wang ◽  
Xiaoying Rong ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Dna Sequences ◽  
Single Copy ◽  
Repetitive Dna Sequences

The use of double fluorescence in situ hybridization to physically map the positions of 5S rDNA genes in relation to the chromosomal location of 18S–5.8S–26S rDNA and a C genome specific DNA sequence in the genus Avena

Genome ◽  
1996 ◽  
Vol 39 (3) ◽  
pp. 535-542 ◽  
Author(s):  
Concha Linares ◽  
Juan González ◽  
Esther Ferrer ◽  
Araceli Fominaya
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Diploid Species ◽  
5S Rdna ◽  
Rdna Genes ◽  
26S Rdna ◽  
A Genome ◽  
Rdna Loci ◽  
C Genome

A physical map of the locations of the 5S rDNA genes and their relative positions with respect to 18S–5.8S–26S rDNA genes and a C genome specific repetitive DNA sequence was produced for the chromosomes of diploid, tetraploid, and hexaploid oat species using in situ hybridization. The A genome diploid species showed two pairs of rDNA loci and two pairs of 5S loci located on both arms of one pair of satellited chromosomes. The C genome diploid species showed two major pairs and one minor pair of rDNA loci. One pair of subtelocentric chromosomes carried rDNA and 5S loci physically separated on the long arm. The tetraploid species (AACC genomes) arising from these diploid ancestors showed two pairs of rDNA loci and three pairs of 5S loci. Two pairs of rDNA loci and 2 pairs of 5S loci were arranged as in the A genome diploid species. The third pair of 5S loci was located on one pair of A–C translocated chromosomes using simultaneous in situ hybridization with 5S rDNA genes and a C genome specific repetitive DNA sequence. The hexaploid species (AACCDD genomes) showed three pairs of rDNA loci and six pairs of 5S loci. One pair of 5S loci was located on each of two pairs of C–A/D translocated chromosomes. Comparative studies of the physical arrangement of rDNA and 5S loci in polyploid oats and the putative A and C genome progenitor species suggests that A genome diploid species could be the donor of both A and D genomes of polyploid oats. Key words : oats, 5S rDNA genes, 18S–5.8S–26S rDNA genes, C genome specific repetitive DNA sequence, in situ hybridization, genome evolution.

Conserved repetitive DNA sequences (Bkm) in normal equine males and sex-reversed females detected by in situ hybridization

1988 ◽  
Vol 48 (2) ◽  
pp. 99-102 ◽  
Author(s):  
M.G. Kent ◽  
K.O. Elliston ◽  
W. Shroeder ◽  
K.S. Guise ◽  
S.S. Wachtel
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Dna ◽  
Dna Sequences ◽  
Repetitive Dna Sequences

scholarly journals Detection of Canine Parvovirus in Naturally Infected Dogs with Enteritis and Myocarditis by in Situ Hybridization

1997 ◽  
Vol 9 (3) ◽  
pp. 255-260 ◽  
Author(s):  
Whan-Gook Nho ◽  
Jung-Hyang Sur ◽  
Alan R. Doster ◽  
Soon-Bok Kim
Keyword(s):  
In Situ Hybridization ◽  
Capsid Protein ◽  
Dna Sequences ◽  
Middle Part ◽  
Canine Parvovirus ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Liver And Kidney ◽  
Labeled Probe

An improved method for the diagnosis of canine parvovirus using in situ hybridization in standard formalin-fixed, paraffin-embedded tissue sections was developed. A digoxigenin-labeled probe complementary to DNA sequences that code for the entire sequence of the capsid protein VP-1 and the middle part of the sequence of the capsid protein VP-2 was designed. Specific histologic localization of canine parvovirus-infected cells was demonstrated in small intestine, tonsil, lymph node, thymus, spleen, heart, liver, and kidney from dogs diagnosed at necropsy with canine parvovirus infection. The in situ hybridization accurately pinpointed the specific sites of viral infection. The detection of canine parvovirus in liver, kidney, and heart tissues together in the same pups could represent an enhanced virulence of this strain of canine parvovirus and suggests a broadened tissue tropism not seen before in Korean strains of canine parvovirus.

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