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PMID: 6261235 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Cell-free coupled transcription-translation system for investigation of linear DNA segments.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 77 ·No. 12 ·1980-12-00 ·Pages 7029-33

Yang HL, Ivashkiv L, Chen HZ, Zubay G, Cashel M

Abstract

Heretofore the DNA-directed coupled transcription-translation system, most useful in gene expression analysis, has been limited to the use of circular or long linear DNAs. Linear DNAs are degraded in this system by an exonucleolytic activity that can be eliminated by making the synthetic extracts from a suitable recB mutant of Escherichia coli. Using these extracts, we have examined the gene expression of a variety of linear DNAs. In particular, the complex pattern of expression of ribosomal protein genes and RNA polymerase genes in the rpoBC-rplLJ region has been analyzed by comparing the protein products obtained when using lambda rifd18 DNA with the product obtained when using the same DNA segmented with various restriction enzymes. The results obtained confirm the conclusions of others obtained by much more elaborate in vivo techniques. It seems highly likely that this cell-free system will have extensive applications in the area of analysis of gene expression.

MeSH Terms
Base Sequence Cell-Free System DNA, Bacterial/genetics DNA, Circular/genetics DNA, Superhelical/genetics Escherichia coli/genetics Exonucleases/metabolism Plasmids Protein Biosynthesis Recombination, Genetic Templates, Genetic Transcription, Genetic
Chemicals
DNA, Bacterial DNA, Circular DNA, Superhelical Exonucleases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Yang H L
Ivashkiv L
Chen H Z
Zubay G
Cashel M
References (27)
27 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.77.12.7029
Published
1980-12-00
Pages
7029-33
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC350434
Subset
IM
Grants
PHS HHS · A II 3277-03 · United States
NIGMS NIH HHS · GML6648-12 · United States
External Links
PubMed source DOI: 10.1073/pnas.77.12.7029 PMC Full Text View journal details
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