MOLECULAR CLONING OF α-AMYLASE GENES FROM DROSOPHILA MELANOGASTER. I. CLONE ISOLATION BY USE OF A MOUSE PROBE

Genetics ◽  
1985 ◽  
Vol 110 (2) ◽  
pp. 299-312
Author(s):  
Robert M Gemmill ◽  
Jack N Levy ◽  
Winifred W Doane
Keyword(s):  
Drosophila Melanogaster ◽  
Dna Sequences ◽  
Genomic Dna ◽  
Cdna Sequence ◽  
Lambda Phage ◽  
Hybridization Probe ◽  
Class A ◽  
Class B ◽  
Small Set

ABSTRACT A cloned ä-amylase cDNA sequence from the mouse is homologous to a small set of DNA sequences from Drosophila melanogaster under appropriate conditions of hybridization. A number of recombinant lambda phage that carry homologous Drosophila genomic DNA sequences were isolated using the mouse clone as a hybridization probe. Putative amylase clones hybridized in situ to one or the other of two distinct sites in polytene chromosome 2R and were assigned to one of two classes, A and B. Clone λDm32, representing class A, hybridizes within chromosome section 53CD. Clone λDm65 of class B hybridizes within section 54A1-B1. Clone λDm65 is homologous to a 1450- to 1500-nucleotide RNA species, which is sufficiently long to code for α-amylase. No RNA homologous to λDm32 was detected. We suggest that the class B clone, λDm65, contains the functional Amy structural gene(s) and that class A clones contain an amylase pseudogene.

Molecular cytogenetic analysis of durum wheat × tritordeum hybrids

Genome ◽  
1997 ◽  
Vol 40 (3) ◽  
pp. 362-369 ◽  
Author(s):  
J. Lima-Brito ◽  
H. Guedes-Pinto ◽  
G. E. Harrison ◽  
J. S. Heslop-Harrison
Keyword(s):  
In Situ Hybridization ◽  
Plant Breeding ◽  
Durum Wheat ◽  
Dna Sequences ◽  
Genomic Dna ◽  
Tandem Repeats ◽  
Nuclear Architecture ◽  
Molecular Cytogenetics ◽  
Hordeum Chilense

Southern and in situ hybridization were used to examine the chromosome constitution, genomic relationships, repetitive DNA sequences, and nuclear architecture in durum wheat × tritordeum hybrids (2n = 5x = 35), where tritordeum is the fertile amphiploid (2n = 6x = 42) between Hordeum chilense and durum wheat. Using in situ hybridization, H. chilense total genomic DNA hybridized strongly to the H. chilense chromosomes and weakly to the wheat chromosomes, which showed some strongly labelled bands. pHcKB6, a cloned repetitive sequence isolated from H. chilense, enabled the unequivocal identification of each H. chilense chromosome at metaphase. Analysis of chromosome disposition in prophase nuclei, using the same probes, showed that the chromosomes of H. chilense origin were in individual domains with only limited intermixing with chromosomes of wheat origin. Six major sites of 18S–26S rDNA genes were detected on the chromosomes of the hybrids. Hybridization to Southern transfers of restriction enzyme digests using genomic DNA showed some variants of tandem repeats, perhaps owing to methylation. Both techniques gave complementary information, extending that available from phenotypic, chromosome morphology, or isozyme analysis, and perhaps are useful for following chromosomes or chromosome segments during further crossing of the lines in plant breeding programs.Key words: In situ hybridization, molecular cytogenetics, plant breeding, Hordeum chilense, Southern hybridization, durum wheat, hybrids.

Detection of maize DNA sequences amplified in wheat

Genome ◽  
1995 ◽  
Vol 38 (5) ◽  
pp. 946-950 ◽  
Author(s):  
Juan Zhang ◽  
Bernd Friebe ◽  
Bikram S. Gill
Keyword(s):  
Triticum Aestivum ◽  
Zea Mays ◽  
In Situ Hybridization ◽  
Dna Sequences ◽  
Hexaploid Wheat ◽  
Chinese Spring ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Metaphase Chromosomes

Genomic in situ hybridization to somatic metaphase chromosomes of hexaploid wheat cv. Chinese Spring using biotinylated maize genomic DNA as a probe revealed the existence of amplified maize DNA sequences in five pairs of chromosomes. The in situ hybridization sites were located on chromosomes 1A, 7A, 2B, 3B, and 7B. One pair of in situ hybridization sites was also observed in hexaploid oat. The locations and sizes of in situ hybridization sites varied among progenitor species.Key words: Triticum aestivum, Zea mays, shared DNA sequences, genomic in situ hybridization.

scholarly journals The Nature of Zeolite Catalytic Sites

2021 ◽  
Author(s):  
◽  
Linda Margaret Parker
Keyword(s):  
Mass Spectrometry ◽  
Carboxylic Acids ◽  
In Situ Ftir ◽  
Electron Pairs ◽  
Desorption Mass Spectrometry ◽  
Class A ◽  
Ftir Studies ◽  
Catalytic Sites ◽  
Class B

<p>The nature of zeolite catalytic sites was studied by observing their interactions with sorbants. In situ FTIR studies of a range of sorbants on H+ZSM-5, H+mordenite and H+Y showed that the zeolite Bronsted proton was transferred towards the sorbant. Sorbants could be placed in three classes depending on the type of hydrogen bond formed. "Class A" sorbants were alcohols, alkanes, ammonia, amines and carboxylic acids and showed a single v0-H band shifted from the vO-H of the zeolite. The shift in vO-H increased with increasing proton affinity of the sorbant. "Class B" sorbants were alkenes and aromatics and showed a resultant broad, flat vO-H due to bonding through the t electrons of the double bond or aromatic ring. "Class C" sorbants included water, ethers, ketenes, aldehydes, nitriles and carboxylic acids (also Class A). They showed extremely broad hydroxyl bands from -3700 to -1200 cm-1 with several maxima. Bonding was through oxygen or nitrogen lone electron pairs. A novel, low temperature (-400 [degrees] C), reaction of acetic acid to ketene was observed over alkali-exchanged zeolites. Thermal desorption/mass spectrometry, mini-reactor mass spectrometry and in situ FTIR techniques were used to investigate the products obtained by varying the carboxylic acid and the catalyst, and the reaction mechanism.</p>

scholarly journals 5-Methylcytosine distribution and genome organization in triticale before and after treatment with 5-azacytidine

1999 ◽  
Vol 112 (23) ◽  
pp. 4397-4404 ◽  
Author(s):  
A. Castilho ◽  
N. Neves ◽  
M. Rufini-Castiglione ◽  
W. Viegas ◽  
J.S. Heslop-Harrison
Keyword(s):  
In Situ Hybridization ◽  
Restriction Fragment ◽  
Dna Sequences ◽  
Genomic Dna ◽  
Restriction Enzymes ◽  
Hybrid Plant ◽  
Rrna Genes ◽  
Next Generation ◽  
Root Tips

Triticale (2n=6x=42) is a hybrid plant including rye (R) and wheat (A and B) genomes. Using genomic in situ hybridization with rye DNA as a probe, we found the chromosomes of the R genome were not intermixed with the wheat chromosomes in 85% of nuclei. After treatment of seedlings with low doses of the drug 5-azacytidine (5-AC), leading to hypomethylation of the DNA, the chromosomes became intermixed in 60% of nuclei; the next generation showed intermediate organization. These results correlate with previous data showing that expression of R-genome rRNA genes, normally suppressed, is activated by 5-AC treatment and remains partially activated in the next generation. The distribution of 5-methylcytosine (5-mC) was studied using an antibody to 5-mC. Methylation was detected along the lengths of all chromosomes; there were some chromosome regions with enhanced and reduced methylation, but these were not located at consistent positions, nor were there differences between R and wheat genome chromosomes. After 5-AC treatment, lower levels of methylation were detected. After 5-AC treatment, in situ hybridization with rye genomic DNA sometimes showed micronuclei of rye origin and multiple translocations between wheat and rye chromosomes. Genomic DNA was analysed using methylation-sensitive restriction enzymes and, as probes, two rDNA sequences, two tandemly organised DNA sequences from rye (pSc200 and pSc250), and copia and the gypsy group retrotransposon fragments from rye and wheat. DNA extracted immediately after 5-AC treatment was cut more by methylation-sensitive restriction enzymes than DNA from untreated seedlings. Each probe gave a characteristic restriction fragment pattern, but rye- and wheat-origin probes behaved similarly, indicating that hypomethylation was induced in both genomes. In DNA samples from leaves taken 13–41 days after treatment, RFLP (Restriction Fragment Length Polymorphism) patterns were indistinguishable from controls and 5-AC treatments with all probes. Surprising differences in hybridization patterns were seen between DNA from root tips and leaves with the copia-fragment probes.

scholarly journals Fixation of transposable elements in the Drosophila melanogaster genome

2005 ◽  
Vol 85 (3) ◽  
pp. 195-203 ◽  
Author(s):  
XULIO MASIDE ◽  
STAVROULA ASSIMACOPOULOS ◽  
BRIAN CHARLESWORTH
Keyword(s):  
Drosophila Melanogaster ◽  
Transposable Elements ◽  
Genomic Dna ◽  
Molecular Level ◽  
Hybridization Technique ◽  
Small Interfering Rnas ◽  
Heterochromatin Formation ◽  
Large Clusters

We have investigated at the molecular level four cases in which D. melanogaster middle repetitive DNA probes consistently hybridized to a particular band on chromosomes sampled from a D. melanogaster natural population. Two corresponded to true fixations of a roo and a Stalker element, and the others were artefacts of the in situ hybridization technique caused by the presence of genomic DNA flanking the transposable elements (TEs) in the probes. The two fixed elements are located in the β-heterochromatin (20A and 80B, respectively) and are embedded in large clusters of other elements, many of which may also be fixed. We also found evidence that this accumulation is an ongoing process. These results support the hypothesis that TEs accumulate in the non-recombining part of the genome. Their implications for the effects of TEs on determining the chromatin structure of the host genomes are discussed in the light of recent evidence for the role of TE-derived small interfering-RNAs as cis-acting determinants of heterochromatin formation.

scholarly journals Tumorous-head (tuh-1; tuh-3) modulates Abd-B bithorax-complex functions in Drosophila melanogaster.

Genetics ◽  
1993 ◽  
Vol 133 (3) ◽  
pp. 593-604
Author(s):  
D T Kuhn ◽  
J A Mack ◽  
C Duan ◽  
G Packert
Keyword(s):  
Maternal Effect ◽  
Bithorax Complex ◽  
Class A ◽  
Complex Functions ◽  
Class B ◽  
In Trans ◽  
Class C ◽  
Specific Cluster ◽  
R Functions

Abstract Abdominal-B (Abd-B) mutants of the bithorax-complex (BX-C) were studied in trans with tuh-3 to evaluate their interactions with this homeotic mutant and the maternal effect locus (tuh-1) controlling tuh-3 expression. Head defects occur in tuh-3 offspring from tuh-1h homozygous mothers, while genital defects occur in homozygous tuh-3 offspring from mothers carrying the tuh-1g allele. The influence exerted by the tuh-1 maternal effects on tuh-3 Abd-B transcript distribution was evaluated by whole mount in situ hybridization. Results demonstrated that: (1) of the 21 Abd-B mutants tested, head defects were produced by SGA62, I127B, I127O and tuh-3, with I127B and tuh-3 as the only mutants to require the head defect maternal effect for expression; (2) one specific cluster of regulatory (r) mutants, Uab1, 65 and I127B, enhanced penetrance and expressivity of tuh-3 head defects; (3) the most profound suppression of head defect penetrance occurred when Abd-B mutants eliminated the morphogenetic (m) and r functions; (4) genital defects increased in frequency in tuh-3/Abd-B mutant trans-heterozygotes with loss of r function; (5) Abd-B transcription (class A, class B, class C) appears normal in tuh-3 embryos when their mothers pass on the tuh-1h head defect maternal effect, whereas the regulatory transcripts (class B and class C) are reduced when tuh-3 mothers pass on the tuh-1g genital disc maternal effect; (5) tuh eye-antennal imaginal discs express ABD-B protein; and (6) tuh-3 misregulates both m and r function of Abd-B.

scholarly journals The Nature of Zeolite Catalytic Sites

2021 ◽  
Author(s):  
◽  
Linda Margaret Parker
Keyword(s):  
Mass Spectrometry ◽  
Carboxylic Acids ◽  
In Situ Ftir ◽  
Electron Pairs ◽  
Desorption Mass Spectrometry ◽  
Class A ◽  
Ftir Studies ◽  
Catalytic Sites ◽  
Class B

<p>The nature of zeolite catalytic sites was studied by observing their interactions with sorbants. In situ FTIR studies of a range of sorbants on H+ZSM-5, H+mordenite and H+Y showed that the zeolite Bronsted proton was transferred towards the sorbant. Sorbants could be placed in three classes depending on the type of hydrogen bond formed. "Class A" sorbants were alcohols, alkanes, ammonia, amines and carboxylic acids and showed a single v0-H band shifted from the vO-H of the zeolite. The shift in vO-H increased with increasing proton affinity of the sorbant. "Class B" sorbants were alkenes and aromatics and showed a resultant broad, flat vO-H due to bonding through the t electrons of the double bond or aromatic ring. "Class C" sorbants included water, ethers, ketenes, aldehydes, nitriles and carboxylic acids (also Class A). They showed extremely broad hydroxyl bands from -3700 to -1200 cm-1 with several maxima. Bonding was through oxygen or nitrogen lone electron pairs. A novel, low temperature (-400 [degrees] C), reaction of acetic acid to ketene was observed over alkali-exchanged zeolites. Thermal desorption/mass spectrometry, mini-reactor mass spectrometry and in situ FTIR techniques were used to investigate the products obtained by varying the carboxylic acid and the catalyst, and the reaction mechanism.</p>

Isolation, characterization, and analysis of Leymus-specific DNA sequences

Genome ◽  
2003 ◽  
Vol 46 (4) ◽  
pp. 673-682 ◽  
Author(s):  
Sigridur Klara Bödvarsdóttir ◽  
Kesara Anamthawat-Jónsson
Keyword(s):  
Dna Sequences ◽  
Genomic Dna ◽  
Southern Hybridization ◽  
Genetic Relatedness ◽  
Repetitive Sequences ◽  
Nucleolar Organiser Regions ◽  
Basic Genome ◽  
Specific Sequences ◽  
Leymus Mollis

Genomic Southern hybridization using labeled total genomic DNA of Leymus mollis as probe showed intense hybridization signals on all restriction enzyme digested DNA from five species of Leymus Hochst., and four species of Psathyrostachys Nevski. Experiments using the same L. mollis probe, but with unlabeled blocking DNA from Psathyrostachys, showed no hybridization at all. These two genera evidently had the same genomic content. Southern hybridization without blocking allowed identification of DNA fragments abundant in Leymus and Psathyrostachys. Fragments potentially specific to Leymus were cloned. Five repetitive DNA clones from L. mollis and L. arenarius were characterized: pLmIs1, pLmIs44, pLmIs51, pLmIs53, and pLaIs56. These clones hybridized to both Leymus and Psathyrostachys on Southern blots — no clone hybridized to only one of these genera. Both Southern blot and fluorescence in situ hybridization (FISH) experiments showed that all the clones contained dispersed repetitive sequences. They painted all and whole chromosomes uniformly except at centromeres, telomeres, and nucleolar organiser regions. Three of these clones, i.e., pLmIs1, pLmIs44, and pLmIs53, were essentially specific to Leymus and Psathyrostachys — little or no hybridization was detected in other genera such as Triticum, Hordeum, Thinopyrum, or Elymus. Sequence analysis further revealed that the clones were part of retroelements. In particular, the clone pLmIs44 produced hybridization profiles suitable for analysis of genetic relatedness among species. The present study shows that Leymus and Psathyrostachys share the same basic genome, Ns, and therefore provides strong evidence for combining these two genera.Key words: Triticeae, Leymus, Psathyrostachys, genome-specific sequences, retrotransposons.

Isolation of A/D and C genome specific dispersed and clustered repetitive DNA sequences from Avena sativa

Genome ◽  
2002 ◽  
Vol 45 (2) ◽  
pp. 431-441 ◽  
Author(s):  
Evgueni V Ananiev ◽  
M Isabel Vales ◽  
Ronald L Phillips ◽  
Howard W Rines
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Dna ◽  
Dna Sequences ◽  
Avena Sativa ◽  
Genomic Dna ◽  
Repetitive Sequences ◽  
Repeated Sequences ◽  
C Genome ◽  
Copy Dna

DNA gel-blot and in situ hybridization with genome-specific repeated sequences have proven to be valuable tools in analyzing genome structure and relationships in species with complex allopolyploid genomes such as hexaploid oat (Avena sativa L., 2n = 6x = 42; AACCDD genome). In this report, we describe a systematic approach for isolating genome-, chromosome-, and region-specific repeated and low-copy DNA sequences from oat that can presumably be applied to any complex genome species. Genome-specific DNA sequences were first identified in a random set of A. sativa genomic DNA cosmid clones by gel-blot hybridization using labeled genomic DNA from different Avena species. Because no repetitive sequences were identified that could distinguish between the A and D gneomes, sequences specific to these two genomes are refereed to as A/D genome specific. A/D or C genome specific DNA subfragments were used as screening probes to identify additional genome-specific cosmid clones in the A. sativa genomic library. We identified clustered and dispersed repetitive DNA elements for the A/D and C genomes that could be used as cytogenetic markers for discrimination of the various oat chromosomes. Some analyzed cosmids appeared to be composed entirely of genome-specific elements, whereas others represented regions with genome- and non-specific repeated sequences with interspersed low-copy DNA sequences. Thus, genome-specific hybridization analysis of restriction digests of random and selected A. sativa cosmids also provides insight into the sequence organization of the oat genome.Key words: oat, cosmid library, in situ hybridization.

Molecular cytogenetic analysis of DNA sequences with flanking telomeric repeats inTriticum aestivumcv. Begra

Genome ◽  
2001 ◽  
Vol 44 (1) ◽  
pp. 133-136
Author(s):  
Marta Dobrzanska ◽  
Elzbieta Kraszewska ◽  
Maria Bucholc ◽  
Glyn Jenkins
Keyword(s):  
Triticum Aestivum ◽  
Dna Sequences ◽  
Genomic Dna ◽  
Repeat Unit ◽  
Chromosome Complement ◽  
Mobile Dna ◽  
Chromosomal Distribution ◽  
Telomeric Sequences ◽  
Dna Fragment

A cloned genomic DNA fragment (pTa241) formerly derived from a DNA fraction obtained from isolated nuclei of embryos of a Polish cultivar of wheat (Triticum aestivum cv. Begra) comprises a tandem repeat of the telomeric array CCCTAAA, and hybridizes in situ exclusively to the telomeres of all chromosome arms of the somatic chromosome complement of wheat. A second cloned fragment (pTa637) derived from the same fraction is 637 bp long, flanked by 28 bp of the same telomeric repeat unit, and hybridizes in situ to the entire lengths of all the chromosomes of the complement. The same pattern of hybridization was observed when the flanking telomeric sequences were removed. A third DNA fragment (pTa1439), derived from unfractionated genomic DNA and flanked with 62 bp of the same telomeric unit, showed the same patterns of distribution. Together with additional evidence from Southern analysis, these observations were interpreted to mean that these sequences are associated with mobile DNA elements and are distributed widely throughout the genome. The chromosomal distribution of the non-telomeric parts of the clones is consistent with the dispersed genomic distribution characteristic of transposons and retroelements.Key words: wheat, Triticum aestivum cv. Begra, mobile elements, telomeric DNA sequence, FISH.

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