Literature DB >> 8041711

Snapshot blotting: transfer of nucleic acids and nucleoprotein complexes from electrophoresis gels to grids for electron microscopy.

S D Jett1, D G Bear.   

Abstract

We present a technique, "snapshot blotting," for the electrophoretic transfer of nucleic acids and nucleoprotein complexes in gel electrophoresis bands onto highly stable carbon film-coated grids for imaging by electron microscopy. The method permits structural analysis of macromolecular species that have been resolved by a gel mobility-shift assay. To demonstrate the efficiency and integrity of the transfer process for a multiprotein-DNA assembly, we have imaged various species of a prokaryotic transcription complex, using the cleavage-defective EcoRI(Q111) protein as an orientation marker and as a blockade of transcription elongation. Snapshot blotting should be of great utility in the structural characterization of nucleic acids and protein-nucleic acid interactions.

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Year:  1994        PMID: 8041711      PMCID: PMC44299          DOI: 10.1073/pnas.91.15.6870

Source DB:  PubMed          Journal:  Proc Natl Acad Sci U S A        ISSN: 0027-8424            Impact factor:   11.205


  16 in total

1.  Studies of DNA-protein interactions by gel electrophoresis.

Authors:  J A Ceglarek; A Revzin
Journal:  Electrophoresis       Date:  1989 May-Jun       Impact factor: 3.535

2.  Measurement of protein-DNA interaction parameters by electrophoresis mobility shift assay.

Authors:  M G Fried
Journal:  Electrophoresis       Date:  1989 May-Jun       Impact factor: 3.535

3.  Glu-111 is required for activation of the DNA cleavage center of EcoRI endonuclease.

Authors:  K King; S J Benkovic; P Modrich
Journal:  J Biol Chem       Date:  1989-07-15       Impact factor: 5.157

4.  The uvsX protein of bacteriophage T4 arranges single-stranded and double-stranded DNA into similar helical nucleoprotein filaments.

Authors:  J Griffith; T Formosa
Journal:  J Biol Chem       Date:  1985-04-10       Impact factor: 5.157

5.  Electron microscopic visualization of DNA in association with cellular components.

Authors:  J D Griffith
Journal:  Methods Cell Biol       Date:  1973       Impact factor: 1.441

6.  Isolation of gram quantities of EcoRI restriction and modification enzymes from an overproducing strain.

Authors:  S C Cheng; R Kim; K King; S H Kim; P Modrich
Journal:  J Biol Chem       Date:  1984-09-25       Impact factor: 5.157

Review 7.  Electron microscope visualization of chromatin and other DNA-protein complexes.

Authors:  J D Griffith; G Christiansen
Journal:  Annu Rev Biophys Bioeng       Date:  1978

8.  Structure of the DNA-Eco RI endonuclease recognition complex at 3 A resolution.

Authors:  J A McClarin; C A Frederick; B C Wang; P Greene; H W Boyer; J Grable; J M Rosenberg
Journal:  Science       Date:  1986-12-19       Impact factor: 47.728

9.  Purification and properties of the sigma subunit of Escherichia coli DNA-dependent RNA polymerase.

Authors:  P A Lowe; D A Hager; R R Burgess
Journal:  Biochemistry       Date:  1979-04-03       Impact factor: 3.162

Review 10.  Use of gel retardation to analyze protein-nucleic acid interactions.

Authors:  D Lane; P Prentki; M Chandler
Journal:  Microbiol Rev       Date:  1992-12
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  1 in total

1.  Electrophoretic mobility shift assay (EMSA) for detecting protein-nucleic acid interactions.

Authors:  Lance M Hellman; Michael G Fried
Journal:  Nat Protoc       Date:  2007       Impact factor: 13.491

  1 in total

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