Toward the construction of a complete physical map of human chromosome 7, we have localized 725 YAC clones to cytogenetically defined regions using fluorescence in situ hybridization (FISH) and by screening with DNA markers of known chromosomal locations. These chromosome 7-specific YAC clones are part of a library constructed with DNA isolated from monochromosomal 7 human-hamster somatic cell hybrid lines. The FISH mapping for 575 clones was accomplished by using "Alu-PCR" amplified YAC DNA against metaphase chromosome spreads made from a monochromosomal 7 human-mouse somatic cell hybrid line. Hybridization- or PCR-based screening of previously mapped DNA markers was performed for the mapping of 221 YAC clones. There was excellent correlation between the map locations obtained for the 71 YACs localized with both methods. All of the regionally localized YAC clones are valuable reagents for mapping and identification of disease genes on human chromosome 7.
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http://dx.doi.org/10.1006/geno.1994.1407 | DOI Listing |
Bio Protoc
January 2025
Biochemistry Department, Western University, London, Canada.
Chloroplast genomes present an alternative strategy for large-scale engineering of photosynthetic eukaryotes. Prior to our work, the chloroplast genomes of (204 kb) and (140 kb) had been cloned using bacterial and yeast artificial chromosome (BAC/YAC) libraries, respectively. These methods lack design flexibility as they are reliant upon the random capture of genomic fragments during BAC/YAC library creation; additionally, both demonstrated a low efficiency (≤ 10%) for correct assembly of the genome in yeast.
View Article and Find Full Text PDFCell Rep Methods
February 2024
Leibniz Institute of Virology (LIV), Hamburg, Germany. Electronic address:
Herpesviruses are large DNA viruses and include important human and veterinary pathogens. Their genomes can be cloned as bacterial artificial chromosomes (BACs) and genetically engineered in Escherichia coli using BAC recombineering methods. While the recombineering methods are efficient, the initial BAC-cloning step remains laborious.
View Article and Find Full Text PDFAntiviral Res
February 2023
Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA; Institute for Drug Discovery, University of Texas Medical Branch, Galveston, TX, USA; Institute for Human Infection and Immunity, University of Texas Medical Branch, Galveston, TX, USA; Institute for Translational Sciences, University of Texas Medical Branch, Galveston, TX, USA; Sealy Institute for Vaccine Sciences, University of Texas Medical Branch, Galveston, TX, USA; Sealy Center for Structural Biology & Molecular Biophysics, University of Texas Medical Branch, Galveston, TX, USA. Electronic address:
Reverse genetic systems are widely used to engineer recombinant viruses with desired mutations. In response to the COVID-19 pandemic, four types of reverse genetic systems have been developed for SARS-CoV-2: (i) a full-length infectious clone that can be used to prepare recombinant SARS-CoV-2 at biosafety level 3 (BSL3), (ii) a trans-complementation system that can be used to produce single-round infectious SARS-CoV-2 at BSL2, (iii) an attenuated SARS-CoV-2 vaccine candidate (with deletions of viral accessory genes) that may be developed for veterinary use as well as for antiviral screening at BSL2, and (iv) replicon systems with deletions of viral structural genes that can be used at BSL2. Each of these genetic systems has its advantages and disadvantages that can be used to address different questions for basic and translational research.
View Article and Find Full Text PDFBiochem J
June 2022
Department of Pathology, Saint Louis University School of Medicine, St. Louis, MO, U.S.A.
Uridine-cytidine kinase like-1 (UCKL-1) is a largely uncharacterized protein with high sequence similarity to other uridine-cytidine kinases (UCKs). UCKs play an important role in the pyrimidine salvage pathway, catalyzing the phosphorylation of uridine and cytidine to UMP and CMP, respectively. Only two human UCKs have been identified, UCK1 and UCK2.
View Article and Find Full Text PDFAntiviral Res
January 2022
College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, Jiangsu, China; Comparative Medicine Research Institute, Yangzhou University, Yangzhou, 225009, Jiangsu, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, Jiangsu, China. Electronic address:
Japanese encephalitis virus (JEV) belongs to a zoonotic flavivirus and is the main cause of viral encephalitis in humans throughout Asia. During the past two decades, both genotypes I (G) and III (GIII) JEVs co-existed in many Asian countries, and the prevalent strains have shifted from GIII to GI. Since all licensed JE vaccines are derived from GIII strains and no specific treatment is available, the development of novel vaccines and specific antiviral drugs against both genotypes is urgently required.
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