Abstract
Several plasmids were constructed in which the SV40 small-t antigen gene was inserted in close proximity downstream from the thermoinducible leftward promoter ( p l ) of bacteriophage λ. Upon temperature induction the best of our constructions expressed a small-t-related 19000-dalton polypeptide in an amount corresponding to approx. 2.5% of total de novo protein synthesis. This 19000-dalton protein was identified as small-t by specific immunoprecipitation with anti-T serum and by two-dimensional fingerprint analysis. In addition to the 19 000-dalton product, representative plasmids expressed fairly large amounts (up to 7% of total de novo protein synthesis) of a protein with an apparent M r of 14500. This 14500-dalton polypeptide was shown to be related to authentic small-t. Presumably the secondary structure of the mRNA starting at is such that translation initiation at an internal AUG codon of the small-t gene is favored over initiation at the true initiating codon.
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