Nucleotide sequence of the simian virus 40 HindII + III restriction fragment D and the total amino acid sequence of the late proteins VP2 and VP3.
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Biomedical subjects
Publications and source records attributed to M Ysebaert.
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The determination of the total 5,224 base-pair DNA sequence of the virus SV40 has enabled us to locate precisely the known genes on the genome. At least 15.2% of the genome is presumably not translated into polypeptides. Particular points of interest revealed by the complete sequence are the initiation of the early t and T antigens at the same position and the fact that the T antigen is coded by two non-contiguous regions of the genome; the T antigen mRNA is spliced in the coding region. In the late region the gene for the major protein VP1 overlaps those for proteins VP2 and VP3 over 122 nucleotides but is read in a different frame. The almost complete amino acid sequences of the two early proteins as well as those of the late proteins have been deduced from the nucleotide sequence. The mRNAs for the latter three proteins are presumably spliced out of a common primary RNA transcript. The use of degenerate codons is decidedly non-random, but is similar for the early and late regions. Codons of the type NUC, NCG and CGN are absent or very rare.
The nucleotide sequence at the beginning of the restriction fragment Hind-D from Simian virus 40 DNA has been derived by partial chemical degradation of (5'-32P)-labeled restriction fragments followed by analysis on polyacrylamide gel according to Maxam and Gilbert [Proc. Natl Acad. Sci. U.S.A. 74, 560-564 (1977)]. The sequence reported here is 140 nucleotides long. It contains an ATG codon which presumably corresponds to the initiator codon of the VP2 protein. This codon is preceded by an untranslated region which shows several interesting features, such as an alternation of (dA+dT)-rich and (dG+dC)-rich blocks.
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Bacteriophage MS2 RNA is 3,569 nucleotides long. The nucleotide sequence has been established for the third and last gene, which codes for the replicase protein. A secondary structure model has also been proposed. Biological properties, such as ribosome binding and codon interactions can now be discussed on a molecular basis. As the sequences for the other regions of this RNA have been published already, the complete, primary chemical structure of a viral genome has now been established.
The nucleotide sequence of the two minor SV40 DNA restriction fragments Hind L and Hind M is here reported. For this purpose we used 5'-32p-labeled DNA fragments either partially digested with snake venom diesterase for analysis by the wandering spot method or partially degraded with the base specific reagents dimethylsulphate or hydrazine for direct sequence analysis on gel. In the former procedure the strands were separated before degradation, while in the latter the strands were separated after modification but before the degradation. Due to the presence of several termination codons, the region Hind L - Hind M cannot be translated in a polypeptide. Also, no initiation codons for protein synthesis are present in this region.
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