In vitro methylation of predetermined regions in recombinant DNA constructs.

In vitro methylation of predetermined regions in recombinant DNA constructs. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

In vitro methylation of predetermined regions in recombinant DNA constructs. Abstract Text:

I Van Den Wyngaert,R L Adams,S U Kass,

DNA methylation at position 5 in the cytosine ring in the sequence CpG can be detrimental to the transcription of a variety of genes in higher eukaryotes (1,2). Although the significance of this transcriptional repression is currently under debate (3,4), there is little disagreement that it plays an important role in genomic imprinting and X-chromosome inactivation (5,6). To study the effects of DNA methylation on transcription in an experimental system, bacterial DNA methyltransferases have been used widely in order to mimic the DNA methylation pattern of eukaryotic genes. However, usually every target site in a given recombinant DNA molecule will be subject to DNA methylation by making use of those enzymes. This might result in an exaggeration of the effects of DNA methylation, as most recombinant DNA molecules contain a high degree of prokaryotic DNA, which is rich in CpGs. This methylated CpGrich DNA can contribute to the effects of DNA methylation by formation of a repressive chromatin structure (7,8). In addition, selective DNA methylation is required to distinguish the effects of DNA methylation on transcription initiation and transcript elongation (8,9). Thus, there is a requirement for a method to generate recombinant DNA molecules that are methylated in a predetermined region. The chapter following this one will describe a method that makes use of ligation of methylated DNA fragments into unmethylated vector DNA. This method relies on the availability of suitable restriction sites, which allow directional cloning of the fragment and, in addition, requires a highly efficient ligation reaction.

In vitro methylation of predetermined regions in recombinant DNA constructs. Publishing Authors By Initials

I Van Den Wyngaert,RL Adams,SU Kass,

For similar abstracts research abstracts see: abstracts research

PUBMED ID PMID:

MEDLINE DATE:

In vitro methylation of predetermined regions in recombinant DNA constructs. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Methods in molecular biology (Clifton, N.J.)

VOLUME: 181

Page Numbers: 243-50

Journal Abbreviation: Methods Mol. Biol.

ISSN: 1064-3745

DAY: 4

MONTH: 07

YEAR: 2001

In vitro methylation of predetermined regions in recombinant DNA constructs. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 9214969

In vitro methylation of predetermined regions in recombinant DNA constructs. Keywords Mesh Terms:

KEYWORDS:

MESH TERMS:

Chemical & Substance for Abstract: In vitro methylation of predetermined regions in recombinant DNA constructs. Information

Substance Name:

Registry Number:

Grant and Affiliation Information for In vitro methylation of predetermined regions in recombinant DNA constructs.

AFFILIATION:

Country: United States

AGENCY:

GRANT:

ACRONYM:

MEDLINETA: Methods Mol Biol

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

In vitro methylation of predetermined regions in recombinant DNA constructs Related Publications

• A colorimetric microanalytical method for acetate and fluoroacetate.
• Adrenergic regulation of a key cardiac potassium channel can contribute to atrial fibrillation: evidence from an IKs transgenic mouse.
• Advanced glycation endproduct crosslink breaker (alagebrium) improves endothelial function in patients with isolated systolic hypertension.
• Altered Na+ channels promote pause-induced spontaneous diastolic activity in long QT syndrome type 3 myocytes.
• An epidemic of dyssynchrony: but what does it mean?
• Assessment of the ethical review process in Sudan.
• Comparison of direct immunofluorescence, immunoassays, and fecal flotation for detection of Cryptosporidium spp. and Giardia spp. in naturally exposed cats in 4 Northern California animal shelters.
• Comparison of initial intraocular pressure response with topical beta-adrenergic antagonists and prostaglandin analogues in African American and white individuals in the Ocular Hypertension Treatment Study.
• Comparison of ophthalmic measurements obtained via high-frequency ultrasound imaging in four species of snakes.
• Control of in vivo contraction/relaxation kinetics by myosin binding protein C: protein kinase a phosphorylation dependent and independent regulation.
• Coverage-adjusted entropy estimation.
• Determination of the in vitro susceptibility of feline tritrichomonas foetus to 5 antimicrobial agents.
• Estimation of central pressure augmentation using automated radial artery tonometry.
• Fading sodium channels in failing hearts.
• Hydrogen-deuterium exchange and selective labeling of deprotonated amino acids and peptides in the gas phase.
• Impact of arterial load and loading sequence on left ventricular tissue velocities in humans.
• Impaired chronotropic and vasodilator reserves limit exercise capacity in patients with heart failure and a preserved ejection fraction.
• In vitro methylation of predetermined regions in recombinant DNA constructs.
• Molecular basis of ranolazine block of LQT-3 mutant sodium channels: evidence for site of action.
• Mutation of an A-kinase-anchoring protein causes long-QT syndrome.
• Pathology partnerships that save lives worldwide.
• Recent integrations of mammalian Hmg retropseudogenes.
• Reversing chronic remodeling in heart failure.
• Semi-automated data processing of hydrogen exchange mass spectra using HX-Express.
• Shear stress inhibition of H(2)O (2) induced p66 (Shc) phosphorylation by ASK1-JNK inactivation in endothelium.
• Small-molecule therapies for cardiac hypertrophy: moving beneath the cell surface.
• Therapeutic potential of tetrahydrobiopterin for treating vascular and cardiac disease.
• Validated prediction model for the development of primary open-angle glaucoma in individuals with ocular hypertension.
• Wechsler's mental deterioration index in the diagnosis of organic brain disease.
• Zwitterion-dianion complexes and anion-anion clusters with negative dissociation energies.