Analysis in Cos-1 cells of processing and polyadenylation signals by using derivatives of the herpes simplex virus type 1 thymidine kinase gene.
Open Access
- 1 February 1983
- journal article
- research article
- Published by Taylor & Francis in Molecular and Cellular Biology
- Vol. 3 (2), 267-279
- https://doi.org/10.1128/mcb.3.2.267
Abstract
Bal31 nuclease was used to resect the herpes simplex virus type 1 thymidine kinase (tk) gene from its 3' end, and a plasmid, pTK206, was isolated that lacked the processing and polyadenylation signals normally found at the 3' end of the gene. The wild-type gene, pTK2, and pTK206 were each transferred to pSV010, a plasmid containing the simian virus 40 (SV40) origin of DNA replication, allowing replication and analysis of the patterns of transcription in Cos-1 cells. Fragments of DNA containing processing and polyadenylation signals from SV40 and polyoma virus were inserted into the 3' end of the resected tk gene, pTK206. We found that tk gene expression requires a processing and polyadenylation signal, that signals from SV40 and polyoma virus could substitute for the herpes simplex virus tk signal, and that considerable differences in the levels of tk mRNA were present in Cos-1 cells transfected by these gene constructs. In addition, tk gene expression was restored to a low level after the insertion of an 88-base-pair fragment from the middle of the SV40 early region. Processing and polyadenylation do not occur in the vicinity of this fragment in SV40, even though it contains the hexanucleotide 5'-AAUAAA-3'.This publication has 76 references indexed in Scilit:
- Controls of RNA splicing and termination in the major late adenovirus transcription unitNature, 1981
- Transcription units of adenovirus type 2Journal of Molecular Biology, 1980
- Structure and genomic organization of the mouse dihydrofolate reductase geneCell, 1980
- Coding potential and regulatory signals of the polyoma virus genomeNature, 1980
- The Genome of Simian Virus 40Science, 1978
- Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase IJournal of Molecular Biology, 1977
- Control of breakdown of the polyadenylate sequence in mammalian polyribosomes: role of poly(adenylic acid)-protein interactionsBiochemistry, 1977
- 3′ Non-coding region sequences in eukaryotic messenger RNANature, 1976
- Detection of specific sequences among DNA fragments separated by gel electrophoresisJournal of Molecular Biology, 1975
- Selective extraction of polyoma DNA from infected mouse cell culturesJournal of Molecular Biology, 1967