A subtractive hybridization method to isolate tissue-specific transcripts: application to the selection of new brain-specific products.
A method based on subtractive hybridization of brain complementary DNAs with peripheral messenger RNAs has enabled us to construct an enriched brain-specific cDNA library. Single-stranded cDNAs (ssc DNAs) were synthesized from brain polyadenylated mRNAs and subsequently hybridized with peripheral mRNAs immobilized on nitrocellulose membrane. Unhybridized sscDNAs were converted into double-stranded cDNAs and cloned into plasmid pUC13. The screening of the resulting library showed that a high percentage of the cloned cDNAs corresponded to mRNAs specifically transcribed in the brain.
Maréchal
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D
Laboratoire de Pharmacologie, Université de Liège, Sart Tilman, Belgium.
Forceille
C
C
Breyer
D
D
Delapierre
D
D
Dresse
A
A
D000818:Animals; D001921:Brain / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D005069:Evaluation Studies as Topic; D015723:Gene Library; D008297:Male; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009928:Organ Specificity; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D014158:Transcription, Genetic
D012333:RNA, Messenger; D004247:DNA