By Frank H. Collins, Susan M. Paskewitz (auth.), Kerry F. Harris (eds.)
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The 12H5 fragment shows the HindIII RFLP at the left of the spacer (probably the ETS region), and the 12A fragment shows the EcoRI RFLP at the right end of the spacer, representing the 3' end of 28S and adjacent lOS regions . l. 12 are revealed by probing EcoRI-digested mosquito DNA with the subclone 12A, derived from the 3' end of the 28S coding region. The different restriction fragments detected probably reflect a combination of factors . The fragments in the An. gambiae and An. quadriannulatus lanes are believed to be 5' IGS regions that are relatively conserved within each of these species.
As expected, the medial regions of the spacers tend to vary both within and between species. Preliminary results indicate that small, frequently repeated sequences within these medial spacer regions may prove useful as species-specific, species-diagnostic DNA probes. As with many other eukaryotes, many of the rDNA genes have intervening sequences, some of which appear to be completely species-specific. Our studies thus far suggest a phylogenetic relationship that differs from the relationship inferred from cytogenetic data.
The net effect of this strategy is that most variations in IGS length could be categorized as one of two basic types of fragments: a cluster of subrepeat monomers and monomer multiples resulting from degeneracy of XhoI sites and a comparable array of fragments homologous to the 5' block of IGS DNA. 6. Using this approach to study gene flow among An. gambiae populations in seven different villages in western Kenya, we were able to identify seven different rDNA spacer genotypes based on the presence or absence of XhoI bands of variable length.