ISOLATION OF CANDIDATE GENES FOR MACULAR DEGENERATION USING AN IMPROVED SOLID-PHASE SUBTRACTIVE CLONING TECHNIQUE

An improved solid-phase subtraction procedure was developed to generat e a readily amplifiable library of short cDNA fragments highly enriche d in the macula (target) versus the peripheral region (driver) of the monkey neural retina. The generated clones were sequenced and 63 were analyzed by northern blotting using total RNA from the monkey macula a nd peripheral retina. The results indicate that 32% are highly enriche d in macula, 36% are below the limits of detection and 32% are not enr iched. No clones were found which were enriched in the peripheral reti na. Our technique is therefore successful in identifying novel cDNAs e nriched in the macula area of the neural retina that may represent pot ential candidate genes for hereditary ocular diseases. It should thus be useful in other situations where subtle differences in expression b etween cell types or tissue areas need to be analyzed. (C) 1995 Academ ic Press, Inc.