Fluorescent in situ hybridization (FISH) is a powerful, direct and sensitive technique with a wide resolution range that enables the simultaneous study of multiple targets, labelled in different colours. Spreading techniques, denoted here as 'Fiber-FISH', increase FISH-resolution to the DNA fiber, using decondensed nuclear DNA as hybridization target. FISH could be a powerful analytical tool for thorough physical examination of yeast artificial chromosomes (YACs) which are often chimaeric or contain internal deletions. However, with one exception restricted to meiotic yeast chromosomes, FISH has not been used successfully on yeast/YAC DNA. We have developed a fast and simple method that can be applied routinely for compositional and structural analysis of cosmid and YAC DNA in yeast. It enables precise localization and ordering of clones, resolves overlaps and distances and gives a detailed picture of the integrity and colinearity of both probe and target. The combination of high resolution, signal abundance and short yeast cell cycle allows direct visualization of replicating DNA fibers. In a 400 kb region of the human dystrophin gene, we identified two replication origins, demonstrating that human DNA cloned in yeast is capable of initiating its own replication.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1038/ng0895-477 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Proteomic characterization and comparison of the infective and adult life stage secretomes from Necator americanus and Ancylostoma ceylanicum.
PLoS Negl Trop Dis
January 2025
Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, Australia.
More than 470 million people globally are infected with the hookworms Ancylostoma ceylanicum and Necator americanus, resulting in an annual loss of 2.1 to 4 million disability-adjusted-life-years. Current infection management approaches are limited by modest drug efficacy, the costs associated with frequent mass drug administration campaigns, and the risk of reinfection and burgeoning drug resistance.
View Article and Find Full Text PDFTowards Improved Peptidic α-Ketoamide Inhibitors of the Plasmodial Subtilisin-Like SUB1: Exploration of N-Terminal Extensions and Cyclic Constraints.
ChemMedChem
January 2025
Université de Montpellier: Universite de Montpellier, IBMM, Pôle Chimie Balard, Campus CNRS, 34093, Montpellier, FRANCE.
After more than 15 years of decline, the Malaria epidemy has increased again since 2017, reinforcing the need to identify drug candidates active on new targets involved in at least two biological stages of the Plasmodium life cycle. The SUB1 protease, which is essential for parasite egress in both hepatic and blood stages, would meet these criteria. We previously reported the structure-activity relationship analysis of α-ketoamide-containing inhibitors encompassing positions P4-P2'.
View Article and Find Full Text PDFPerfusion estimation from dynamic non-contrast computed tomography using self-supervised learning and a physics-inspired U-net transformer architecture.
Int J Comput Assist Radiol Surg
January 2025
Department of Biomedical Engineering, The University of Texas at Austin, Austin, TX, USA.
Purpose: Pulmonary perfusion imaging is a key lung health indicator with clinical utility as a diagnostic and treatment planning tool. However, current nuclear medicine modalities face challenges like low spatial resolution and long acquisition times which limit clinical utility to non-emergency settings and often placing extra financial burden on the patient. This study introduces a novel deep learning approach to predict perfusion imaging from non-contrast inhale and exhale computed tomography scans (IE-CT).
View Article and Find Full Text PDFComparison between ZenoTOF 7600 system and QTOF for plant metabolome: an example of metabolomics applied to coffee leaves.
Metabolomics
January 2025
Bioanalysis and Drug Discovery, Faculty of Pharmacy, RD3 Unit of Pharmacognosy, Université Libre de Bruxelles, Bd du Triomphe, Campus Plaine, CP 205/09, 1050, Brussels, Belgium.
Introduction: ZenoTOF is new class of high-resolution mass spectrometer that combines resolution and sensitivity. This mass spectrometer is well designed to perform metabolomics.
Methods: In this context, we compared the performance of ZenoTOF 7600 system (Sciex) with QTOF6520 (Agilent Technologies) through the leaf metabolome analysis of two Coffea species, namely C.
Gas-Phase Fragmentation of Coenzyme Q Radical Anion Generated by APCI: A Study by High/Low-Resolution Tandem/Sequential Mass Spectrometry.
J Am Soc Mass Spectrom
January 2025
Dipartimento di Scienze del Suolo, della Pianta e degli Alimenti, Università degli Studi di Bari Aldo Moro, Via G. Amendola 165/a, 70126 Bari, Italy.
Coenzyme Q (CoQ) and closely related compounds with varying isoprenoid tail lengths (CoQ, = 6-9) are biochemical cofactors involved in many physiological processes, playing important roles in cellular respiration and energy production. Liquid chromatography (LC) coupled with single or tandem mass spectrometry (MS) using electrospray (ESI) or atmospheric pressure chemical ionization (APCI) is considered the gold standard for the identification and quantification of CoQ in food and biological samples. However, the characteristic fragmentation exhibited by the CoQ radical anion ([M], / 862.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!