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. 1980 Nov;36(2):611–616. doi: 10.1128/jvi.36.2.611-616.1980

Construction of a specific amber codon in the simian virus 40 T-antigen gene by site-directed mutagenesis.

D R Rawlins, N Muzyczka
PMCID: PMC353682  PMID: 6253682

Abstract

The site-directed bisulfite mutagenesis technique has been used to construct a specific mutation, am404, at nucleotide position 3124 in the simian virus 40 genome. The mutation was contained within a PstI restriction site (map position 0.27) and prevented cleavage by PstI at that position. Nucleotide sequence analysis of the mutagenized region indicated that only a single base pair change had occurred: a guanosine x cytosine leads to adenine x thymine transition. Comparison of the nucleotide sequence of am404 with the known DNA sequence of simian virus 40 indicted that the mutation in am404 resulted in the conversion of a glutamine codon to an amber codon. am404 could not replicate autonomously when transfected into monkey cells (BSC-40) but did replicate when it was cotransfected with the late deletion helper virus dl1007. On the basis of its position in the T-antigen, gene am404 should produce a T-antigen 24% shorter than the wild-type protein.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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