The significance of certain types of DNA secondary structure was assessed on a purely theoretical basis by analyzing their distribution over the entire genomes of bacteriophages phi X174, G4, fd, the eucaryotic virus SV40 and the E. coli plasmid pBR322. A computer program was designed to search the translated and non-translated (intercistronic) regions of these genomes for the potential of forming perfect hairpins (perfectly base paired stems: no G-T pairing; no interruptions) with small loops. The number of found occurrences was then compared to the statistically expected number. The results reveal a significant excess in each genome of perfect hairpins with stem length greater than 7 and end loops less than or equal to 20 over that expected given their nucleotide composition. For stem length of 5 to 7 the same observation holds true for the intercistronic regions, but for translated regions the number of such structures is below expectation. This suggests an evolutionary selection for these structures.
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