Detection of Trypanosoma cruzi by simplified dot blot with a total genomic DNA probe

1990 ◽  
Vol 18 (5) ◽  
pp. 859-860
Author(s):  
DAVID J. M. LEWIS ◽  
FRANK ASHALL
Keyword(s):  
Trypanosoma Cruzi ◽  
Genomic Dna ◽  
Dna Probe ◽  
Dot Blot

Characterization of a fusion cDNA (RARA/myl) transcribed from the t(15;17) translocation breakpoint in acute promyelocytic leukemia

1992 ◽  
Vol 12 (2) ◽  
pp. 800-810
Author(s):  
K S Chang ◽  
S A Stass ◽  
D T Chu ◽  
L L Deaven ◽  
J M Trujillo ◽  
...  
Keyword(s):  
Cdna Library ◽  
Acute Promyelocytic Leukemia ◽  
Blot Analysis ◽  
Blot Hybridization ◽  
Promyelocytic Leukemia ◽  
Dna Probe ◽  
Translocation Breakpoint ◽  
Chromosome 15 ◽  
Dot Blot ◽  
Mrna Species

A nonrandom chromosomal translocation breakpoint, t(15;17)(q22;q21), is found in almost all patients with acute promyelocytic leukemia (APL). Most of these breakpoints occur within the second intron of the retinoic acid receptor-alpha (RARA) gene. We screened a cDNA library of APL and have identified and sequenced a cDNA transcribed from the t(15;17) translocation breakpoint. The 5' end of cDNA p1715 consists of 503 bp of the RARA exon II sequence. A 1.76-kb cDNA without homology to any known gene available in GenBank was found truncated downstream. This cDNA sequence was assigned to chromosome 15 by dot blot hybridization of the flow cytometry-sorted chromosomes. We designate this fusion cDNA RARA/myl, which is different from myl/RARA reported by de The et al. (H. de The, C. Chomienne, M. Lanotte, L. Degos, and A. Dejean, Nature (London) 347:558-561, 1990). This result demonstrates that the two different types of hybrid mRNA can be transcribed from this breakpoint. We screened a non-APL cDNA library and identified a 2.8-kb myl cDNA. This cDNA is able to encode a polypeptide with a molecular weight of 78,450. Alternative splicing of the myl gene which resulted in myl proteins with different C terminals was found. Southern blot analysis of the genomic DNA isolated from 17 APL patients by using the myl DNA probe demonstrated that the myl gene in 12 samples was rearranged. Northern (RNA) blot analysis of RARA gene expression in two APL RNA samples showed abnormal mRNA species of 4.2 and 3.2 kb in one patient and of 4.8 and 3.8 kb in another patient; these were in addition to the normal mRNA species of 3.7 and 2.7-kb. The myl DNA probe detected a 2.6-kb abnormal mRNA in addition to the normal mRNA species of 3.2, 4.2, and 5.5 kb. Using the polymerase chain reaction, we demonstrated that both RARA/myl and myl/RARA were coexpressed in samples from three different APL patients. From this study, we conclude that the t(15;17) translocation breakpoint results in the transcription of two different fusion transcripts which are expected to be translated into fusion proteins.

scholarly journals Species-Specific Detection of the Maize Pathogens Sporisorium reiliana and Ustilago maydis by Dot Blot Hybridization and PCR-Based Assays

Plant Disease ◽  
1999 ◽  
Vol 83 (4) ◽  
pp. 390-395 ◽  
Author(s):  
M. L. Xu ◽  
A. E. Melchinger ◽  
T. Lübberstedt
Keyword(s):  
Genomic Dna ◽  
Blot Hybridization ◽  
Pathogenic Fungi ◽  
Ustilago Maydis ◽  
Dot Blot ◽  
Cross Hybridization ◽  
Dot Blot Hybridization ◽  
Fungal Dna ◽  
Species Specific ◽  
Sporisorium Reiliana

Head smut of maize, caused by Sporisorium reiliana, may substantially reduce grain yield. The objective of the present study was to develop a highly specific and sensitive DNA-based assay for detection of S. reiliana and its differentiation from Ustilago maydis, a maize fungus inducing the symptomatically similar common smut disease. Plasmid libraries of S. reiliana and U. maydis were constructed using a shotgun cloning procedure. Clones containing strongly hybridizing species-specific DNA were selected by screening libraries with their own labeled genomic DNA, followed by cross-hybridization with genomic DNA of maize and other maize-pathogenic fungi. The selected clones were used to generate subclones with short insert fragments to facilitate PCR amplification for labeling and primer design for a PCR assay. Using Dig-dUTP labeled inserts, detection of less than 0.16 ng of fungal DNA was possible by dot blot hybridization. Sequences of insert fragments were determined to design primer pairs for a PCR-based assay. Primer pairs SR1 and SR3 are species-specific for S. reiliana, and UM11 is species-specific for U. maydis. The PCR-based assays can detect fungal DNA of less than 1.6 pg using SR1 and SR3, and 8 pg using UM11, irrespective of the presence of maize DNA. Use of SR1 and SR3 allowed detection of S. reiliana in the extracts of pith, node, and shank from S. reiliana-infected plants, but not in leaves. Thus, both the dot blot hybridization and the PCR-based assays provide a highly sensitive and reliable tool for detection and differentiation of corn smut caused either by S. reiliana or by U. maydis.

scholarly journals Back Cover: A Mycoplasma Genomic DNA Probe using Gated Nanoporous Anodic Alumina (ChemPlusChem 3/2017)

ChemPlusChem ◽  
2017 ◽  
Vol 82 (3) ◽  
pp. 467-467
Author(s):  
Luís Pla ◽  
Elisabet Xifré-Pérez ◽  
Àngela Ribes ◽  
Elena Aznar ◽  
M. Dolores Marcos ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Anodic Alumina ◽  
Dna Probe ◽  
Back Cover ◽  
Nanoporous Anodic Alumina

scholarly journals Radiolabeled total parasite DNA probe specifically detects Trypanosoma cruzi in mammalian blood.

1988 ◽  
Vol 26 (3) ◽  
pp. 576-578 ◽  
Author(s):  
F Ashall ◽  
D A Yip-Chuck ◽  
A A Luquetti ◽  
M A Miles
Keyword(s):  
Trypanosoma Cruzi ◽  
Dna Probe ◽  
Mammalian Blood

Primers and a Specific DNA Probe for Detecting Lactic Acid Bacteria Producing 3-Hydroxypropionaldehyde from Glycerol in Spoiled Ciders

2001 ◽  
Vol 64 (6) ◽  
pp. 833-837 ◽  
Author(s):  
OLIVIER CLAISSE ◽  
ALINE LONVAUD-FUNEL
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Polymerase Chain Reaction Amplification ◽  
Klebsiella Oxytoca ◽  
Blot Hybridization ◽  
Dna Probe ◽  
Dot Blot ◽  
Glycerol Dehydratase ◽  
Polymerase Chain ◽  
Dna Primers

Of the 40 strains isolated from several spoiled ciders where glycerol was degraded, 36 were identified as Lactobacillus collinoides, three were Lactobacillus hilgardii, and one was Lactobacillus mali. However, only 30 L. collinoides and two L. hilgardii could degrade glycerol. The glycerol dehydratase activity was shown. The main product of the transformation was 1,3 propanediol. Two DNA primers GD1 and GD2 were chosen in the region encoding one of the subunits of glycerol dehydratase of Citrobacter freundii, Klebsiella pneumoniae, Klebsiella oxytoca, Salmonella Typhimurium, and Clostridium pasteurianum. A 279-bp amplicon in polymerase chain reaction amplification was obtained with the genomic L. collinoides IOEB 9527 DNA as template. The amino acid sequence deduced from the amplicon DNA sequence showed a very high similarity and identity with the gene of gram-negative and C. pasteurianum species. After labeling, the amplicon was used as DNA probe in dot-blot hybridization with the genomic DNA of all the tested strains. Only strains that could degrade glycerol hybridized. Moreover, polymerase chain reactions using GD1 and GD2 revealed only glycerol dehydratase genes of positive L. collinoides and L. hilgardii strains. The primers and the amplicon proved to be suitable and reliable tools to detect the lactic acid bacteria involved in the deterioration of cider.

A species-specific oligonucleotide DNA probe for the identification of Meloidogyne incognita

Parasitology ◽  
1991 ◽  
Vol 103 (2) ◽  
pp. 315-319 ◽  
Author(s):  
M. R. Chacon ◽  
R. M. E. Parkhouse ◽  
M. P. Robinson ◽  
P. R. Burrows ◽  
T. Garate
Keyword(s):  
Dna Sequences ◽  
Diagnostic Tool ◽  
Meloidogyne Incognita ◽  
Genomic Dna ◽  
Genomic Library ◽  
Dna Probe ◽  
Equal Length ◽  
Race 1 ◽  
Species Specific

A genomic library of Meloidogyne incognita Race 1 has been prepared in the bacteriophage λgt10 and screened for specific DNA sequences by hybridization with radio-isotope labelled total genomic DNA from a number of Meloidogyne species. One clone isolated (MR1#15), although not totally species specific, clearly showed preferential hybridization to M. incognita. Following subcloning and sequencing of the 255 bp insert, four stretches of the sequence corresponding to oligonucleotides of approximately equal length (~60 bp) were synthesized and examined for specificity. One of them, MR1#15.2, showed the necessary specificity to be used as a diagnostic tool.

scholarly journals Identification of Vibrio halioticoli by colony hybridization with non-radioisotope labeled genomic DNA probe

2002 ◽  
Vol 68 (1) ◽  
pp. 227-229 ◽  
Author(s):  
REIJI TANAKA ◽  
MASASHI OOTSUBO ◽  
TOMOO SAWABE ◽  
KENICHI TAJIMA ◽  
JOHAN VANDENBERGHE ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Dna Probe ◽  
Colony Hybridization
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