Nucleic Acids Res. 1988 Aug 11;16(15):7351-67.

A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions.

Source

Department of Human Genetics, Cetus Corporation, Emeryville, CA 94608.

Abstract

Specific, end-labeled DNA fragments can be simply and rapidly prepared using the polymerase chain reaction (PCR). Such fragments are suitable for use in DNase I protection footprint assays, chemical sequencing reactions, and for the production and analysis of paused RNA polymerase transcription complexes. Moreover, a general means of introducing a specific mutation at any position along the length of such PCR-generated fragments is described. These procedures, which can circumvent the need for large-scale phage or plasmid growths, preparative gel-electrophoresis and the screening of molecular clones, can facilitate the rapid study of sequence-specific interactions of proteins and DNA. A rapid means of removing excess oligonucleotide primers from completed PCRs is also described.

PMID:: 3045756; [PubMed - indexed for MEDLINE]
PMCID:: PMC338413

Free PMC Article

Publication Types, MeSH Terms, Substances, Grant Support

Publication Types

MeSH Terms

Substances

DNA-Directed RNA Polymerases

Grant Support

GM07232/GM/NIGMS NIH HHS/United States

LinkOut - more resources

Full Text Sources

Supplemental Content

Save items

Related citations in PubMed

Footprinting DNA-protein complexes in situ following gel retardation assays using 1,10-phenanthroline-copper ion: Escherichia coli RNA polymerase-lac promoter complexes. [Biochemistry. 1987]
Footprinting DNA-protein complexes in situ following gel retardation assays using 1,10-phenanthroline-copper ion: Escherichia coli RNA polymerase-lac promoter complexes.
Kuwabara MD, Sigman DS. Biochemistry. 1987 Nov 17; 26(23):7234-8.
T4 early promoter strength probed in vivo with unribosylated and ADP-ribosylated Escherichia coli RNA polymerase: a mutation analysis. [Microbiology. 2000]
T4 early promoter strength probed in vivo with unribosylated and ADP-ribosylated Escherichia coli RNA polymerase: a mutation analysis.
Sommer N, Salniene V, Gineikiene E, Nivinskas R, Rüger W. Microbiology. 2000 Oct; 146 ( Pt 10):2643-53.
Characterization of pre-transcription complexes made at a bacteriophage T4 middle promoter: involvement of the T4 MotA activator and the T4 AsiA protein, a sigma 70 binding protein, in the formation of the open complex. [J Mol Biol. 1996]
Characterization of pre-transcription complexes made at a bacteriophage T4 middle promoter: involvement of the T4 MotA activator and the T4 AsiA protein, a sigma 70 binding protein, in the formation of the open complex.
Hinton DM, March-Amegadzie R, Gerber JS, Sharma M. J Mol Biol. 1996 Feb 23; 256(2):235-48.
RNA polymerase-DNA interactions in Streptomyces. In vitro studies of a S. lividans plasmid promoter with S. coelicolor RNA polymerase. [J Mol Biol. 1985]
RNA polymerase-DNA interactions in Streptomyces. In vitro studies of a S. lividans plasmid promoter with S. coelicolor RNA polymerase.
Buttner MJ, Brown NL. J Mol Biol. 1985 Sep 5; 185(1):177-88.
Review Identification of human serum cholinesterase variants using the polymerase chain reaction amplification technique. [Trends Pharmacol Sci. 1989]
Review Identification of human serum cholinesterase variants using the polymerase chain reaction amplification technique.
La Du BN. Trends Pharmacol Sci. 1989 Aug; 10(8):309-13.

See reviews... See all...

Cited by over 100 PubMed Central articles

Lysine decarboxylase catalyzes the first step of quinolizidine alkaloid biosynthesis and coevolved with alkaloid production in leguminosae. [Plant Cell. 2012]
Lysine decarboxylase catalyzes the first step of quinolizidine alkaloid biosynthesis and coevolved with alkaloid production in leguminosae.
Bunsupa S, Katayama K, Ikeura E, Oikawa A, Toyooka K, Saito K, Yamazaki M. Plant Cell. 2012 Mar; 24(3):1202-16. Epub 2012 Mar 13.
Role of position 627 of PB2 and the multibasic cleavage site of the hemagglutinin in the virulence of H5N1 avian influenza virus in chickens and ducks. [PLoS One. 2012]
Role of position 627 of PB2 and the multibasic cleavage site of the hemagglutinin in the virulence of H5N1 avian influenza virus in chickens and ducks.
Schat KA, Bingham J, Butler JM, Chen LM, Lowther S, Crowley TM, Moore RJ, Donis RO, Lowenthal JW. PLoS One. 2012; 7(2):e30960. Epub 2012 Feb 21.
Activation of hepatic lipase expression by oleic acid: possible involvement of USF1. [Nutrients. 2009]
Activation of hepatic lipase expression by oleic acid: possible involvement of USF1.
van Deursen D, van Leeuwen M, Akdogan D, Adams H, Jansen H, Verhoeven AJ. Nutrients. 2009 Feb; 1(2):133-47. Epub 2009 Oct 29.

See all...

Related information

Related Citations
Calculated set of PubMed citations closely related to the selected article(s) retrieved using a word weight algorithm. Related articles are displayed in ranked order from most to least relevant, with the “linked from” citation displayed first.
References for this PMC Article
Citation referenced in PubMed article. Only valid for PubMed citations that are also in PMC.
Free in PMC
Free full text articles in PMC
Cited in PMC
Full-text articles in the PubMed Central Database that cite the current articles.

Recent activity

Clear Turn Off Turn On

A general method of in vitro preparation and specific mutagenesis of DNA fragmen...
A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions.
Nucleic Acids Res. 1988 Aug 11 ;16(15):7351-67.

PubMed
Biomedical versus cultural constructions of abnormality: the case of idiopathic ...
Biomedical versus cultural constructions of abnormality: the case of idiopathic hirsutism in the United States.
Cult Med Psychiatry. 1988 Jun ;12(2):219-38.

PubMed
Glycolipid markers of murine lymphocyte subpopulations.
Glycolipid markers of murine lymphocyte subpopulations.
J Immunol. 1978 Feb ;120(2):676-9.

PubMed
Subpopulations of splenic T cells regulating an antihapten antibody response. I....
Subpopulations of splenic T cells regulating an antihapten antibody response. I. Helper and amplifier cells.
J Immunol. 1978 Feb ;120(2):579-85.

PubMed
Dextromethorphan binding sites in the guinea pig brain.
Dextromethorphan binding sites in the guinea pig brain.
Cell Mol Neurobiol. 1988 Jun ;8(2):149-56.

PubMed

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

PubMed

Result Filters

Display Settings:

Send to: