Large numbers of random point and cluster mutations within the adenovirus VA I gene allow characterization of sequences required for efficient transcription
Open Access
- 1 January 1987
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 15 (20) , 8293-8303
- https://doi.org/10.1093/nar/15.20.8293
Abstract
We have isolated clones with well over 100 randomly dispersed point mutations distributed throughout the 5' half of chemically synthesized adenovirus type 2 VA I genes. In addition, we have isolated clusters of mutations targeted to the regions corresponding to the A and B block consensus sequences of eukaryotic tRNA and adenovirus VA genes. In vitro analyses of these constructs have allowed us to survey in detail the importance of DNA sequence to transcriptional efficiency. Our analyses demonstrate that certain constructs with radically substituted A block regions can be transcribed efficiently. In contrast, there is little tolerance for variation in the sequence within the B block region. We propose that the B block sequence should be R-G-A/T-T-C-R-A-N-N-C for optimal transcriptional efficiency of the VA I gene in mammalian cells.Keywords
This publication has 26 references indexed in Scilit:
- Studies on transformation of Escherichia coli with plasmidsPublished by Elsevier ,2006
- Defining the functional domains in the control region of the adenovirus type 2 specific VARNA1 geneJournal of Molecular Biology, 1987
- A detailed mutational analysis of the eucaryotic tRNA1met gene promoterCell, 1983
- Control region for adenovirus VA RNA transcription.Proceedings of the National Academy of Sciences, 1981
- Transcriptional control regions of the adenovirus VAI RNA geneCell, 1980
- Faithful transcription of adenovirus 5.5 S RNA gene by RNA polymerase III in a human KB cell-free extract.Journal of Biological Chemistry, 1980
- A control region in the center of the 5S RNA gene directs specific initiation of transcription: I. The 5′ border of the regionCell, 1980
- Selective and accurate initiation of transcription at the ad2 major late promotor in a soluble system dependent on purified rna polymerase ii and dnaCell, 1979
- DNA sequencing with chain-terminating inhibitorsProceedings of the National Academy of Sciences, 1977
- RNA of low molecular weight in KB cells infected with adenovirus type 2Journal of Molecular Biology, 1966