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PMID: 270678 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Structural gene identification and mapping by DNA-mRNA hybrid-arrested cell-free translation.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 74 ·No. 10 ·1977-10-00 ·Pages 4370-4

Paterson BM, Roberts BE, Kuff EL

Abstract

We present a simple method for directly correlating structural gene sequences in DNA with their corresponding mRNAs. This is based upon the fact that mRNA hybridized with its complementary DNA will not direct the cell-free synthesis of a complete polypeptide. Full translational activity of the mRNA is recovered upon the heat melting of the hybrid. Utilizing the rabbit beta globin clone PbetaG1, we demonstrate the application of hybrid-arrested translation for the identification of structural gene sequences within recombinant DNA molecules. In addition, the method is used to locate and order precisely several adenovirus 2 polypeptides within the viral genome.

MeSH Terms
Animals Base Sequence Cell-Free System Chromosome Mapping DNA/genetics DNA, Recombinant Genes Nucleic Acid Hybridization Protein Biosynthesis RNA, Messenger/genetics Rabbits
Chemicals
DNA, Recombinant RNA, Messenger DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Paterson B M
Roberts B E
Kuff E L
References (23)
23 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.74.10.4370
Published
1977-10-00
Pages
4370-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC431943
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.74.10.4370 PMC Full Text View journal details
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