Overlap hybridization screening: isolation and characterization of overlapping DNA fragments surrounding the leu2 gene on yeast chromosome III

Gene
A C Chinault, J Carbon

Abstract

A set of four plasmids containing overlapping segments comprising a total of about 30 kbp of cloned DNA from chromosome III of yeast (Saccharomyces cerevisiae) has been isolated and characterized by restriction endonuclease analyses and DNA:DNA hybridizations. Colony hybridization was carried out with labeled pYe(leu2)10, a plasmid carrying the yeast leu2 gene, to a bank of bacterial colonies containing recombinant plasmids constructed from the vector ColE1 and random fragments of yeast DNA. This resulted in the detection of two plasmids, pYe11G4 and pYe40C3, with DNA inserts which partially overlap the original cloned segment and contain additional DNA extending in opposite directions on the chromosome. By carrying out a second round of colony hybridization with pYe40C3, the cloned region was further extended in one direction. A region of DNA that is repeated at least ten times in the yeast genome was identified by hybridization of pYe11G4 to an EcoRI digest of total yeast DNA. The procedure described in this paper should allow the isolation of large sections of chromosomes, including non-transcribed regions, surrounding cloned genes.

References

Jun 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·S Broome, W Gilbert
Feb 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·B Ratzkin, J Carbon
Apr 8, 1977·Science·J S BeckmannJ Abelson
Apr 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·A HinnenG R Fink
Dec 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·A WalzJ Carbon
May 1, 1976·Proceedings of the National Academy of Sciences of the United States of America·K StruhlR W Davis
Oct 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·M Grunstein, D S Hogness
Nov 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·L Clarke, J Carbon
Jun 14, 1967·Journal of Molecular Biology·B Hirt
Aug 15, 1973·Journal of Molecular Biology·P E Lobban, A D Kaiser
Jun 13, 1966·Biochemical and Biophysical Research Communications·D T Denhardt

❮ Previous
Next ❯

Citations

Aug 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·C L Hsiao, J Carbon
Apr 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·L Clarke, J Carbon
Jun 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·C L Hsiao, J Carbon
Mar 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·J BrennandC T Caskey
Jul 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·R L NussbaumC T Caskey
Mar 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·P H YenL J Shapiro
Nov 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·J D NotiA A Szalay
Nov 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·L ClarkeJ Carbon
Apr 24, 1982·Nucleic Acids Research·M J DobsonL A Fothergill
Jan 25, 1983·Nucleic Acids Research·J C Larkin, J L Woolford
Aug 26, 1986·Nucleic Acids Research·J S MillsS M Kingsman
Jun 24, 1988·Nucleic Acids Research·A M FultonA J Kingsman
Aug 1, 1995·Letters in Applied Microbiology·J LacadenaJ G Gavilanes
Sep 17, 2011·Human Mutation·Marion PhylipsenCornelis L Harteveld
Feb 5, 1981·Journal of Molecular Biology·A J KingsmanJ Carbon
Jul 28, 1989·Cell·M BraddockS M Kingsman
Sep 1, 1987·Yeast·M Fitzgerald-Hayes
Jun 21, 2017·International Journal of Genomics·Sonia MayoFrancisco Martínez
Jun 5, 2013·Genetics·David B Kaback

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.