Repeated nucleotide sequence arrays in Balbiani ring 1 of Chironomus tentans contain internally nonrepeating and subrepeating elements.

Balbiani rings in Chironomus are large puffs on salivary gland polytene chromosomes that contain functionally related, but nonidentical genes that code for tissue-specific secretory polypeptides. In situ hybridization was used to select a recombinant plasmid (pCtBR1-1) that contained an insert of Chironomus tentans genomic DNA that originated from Balbiani ring 1. Mapping with restriction endonucleases demonstrated that the insert was 385 bp (base pair) and it contained duplicate clusters of certain cleavage sites about 250 bp apart. This repeat was shown to be part of tandem sequence arrays in the genome by hybridization of radioactive pCtBR1-1 to nitrocellulose blots containing limit and partial restriction endonuclease digests of nuclear DNA. Subsequent sequence analysis of the cloned DNA confirmed the presence of one complete copy of a 246-bp repeat comprised of a 114-bp internally nonrepeating segment and a 132-bp segment containing four 33-bp subrepeats. The subrepeats apparently evolved from a simple 9-bp sequence encoding a consensus tripeptide (Lys-Pro-Ser) in which the first two codons (AAA-CCA) were highly conserved at the nucleotide level. Comparisons between intragenic and interspecific (BRb in Chironomus thummi) copies of corresponding sequences revealed that, during the evolution of these tandemly repeated protein-coding sequences, internally nonrepeated segments were highly conserved and most likely became interspersed by variable segments containing subrepeats that arose from reduplication and divergence of 9-bp repeats.