1887

Abstract

The novel mycobacterial insertion sequence IS was analysed by coupled transcription-translation, of both strands independently, in a cell-free extract using an exogenous promoter. This revealed only one protein product, , as predicted from the nucleotide sequence. The protein was readily translated in recombinant , using the promoter, though it did not appear as a major product by SDS-PAGE analysis. A synthetic peptide was used to generate and affinity-purify a specific anti- antibody, which clearly identified the protein in recombinant . was relatively stable in exponential phase and stationery phase bacteria, though a 28 kDa processed form was seen to accumulate over a period of hours. Both forms appeared in the soluble fraction of the bacterial lysate. The anti- antibody also identified , as a 28 kDa processed product, in Western blots of protein extracts from , indicating a level of expression which would be unusually high for a classical transposase. These data have important implications for the relationship between IS and its host.

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1992-08-01
2021-07-31
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