Abstract

A linked cell-free system has been developed which is capable of transcribing and translating mamalian viral DNA, and its characteristics and requirements are outlined. In this system, simian virus 40 (SV40) DNA Form I (supercoiled) directed the synthesis of discrete polypeptides up to 85,000 daltons in size. One of these products was indistingusihable from authentic major virus capsid protein VPI, as judged by mobility on sodium dodecyl sulfate/polyacrylamide gels, antibody predipitation, and peptide analyses. The cell-free products larger than VPI comprised a number of polypeptides ranging in molecular weight from 50,000 to 85,000. These polypeptides demonstrated no immunological relationship whatsoever to the structural protein VPI. However, two of these products, along with one of approximately 25,000 dlatons, were precipitated with antiserum to SV40 tumor antigen. Linear SV40 DNA generated by the cleavage of Form I DNA with the restriction endonuclease EcoR1 was an efficient template in this system and also directed the synthesis of a polypeptide migrating with VPI on polyacrylamide gels. The potential of this system for defining a functional map of a DNA genome is discussed.

Keywords

CapsidDNARestriction enzymeBiologyMolecular biologySodium dodecyl sulfateEcoRIPolyacrylamide gel electrophoresisVirusBiochemistryVirologyEnzyme

Related Publications

Publication Info

Year
1975
Type
article
Volume
72
Issue
5
Pages
1922-1926
Citations
70
Access
Closed

External Links

View on DOI.org

Social Impact

Altmetric
PlumX Metrics

Social media, news, blog, policy document mentions

Citation Metrics

70
OpenAlex

Cite This

Bryan E. Roberts, Marian Gorecki, Richard C. Mulligan et al. (1975). Simian virus 40 DNA directs synthesis of authentic viral polypeptides in a linked transcription-translation cell-free system.. Proceedings of the National Academy of Sciences , 72 (5) , 1922-1926. https://doi.org/10.1073/pnas.72.5.1922

Identifiers

DOI
10.1073/pnas.72.5.1922