Identifying genes within microdissected genomic DNA: isolation of brain expressed genes from a translocation region associated with inherited mental illness.
An improved protocol has been developed for physical enrichment of cDNA sequences by hybridization to genomic DNA. When applied to microdissection recombinants derived from a translocation breakpoint region associated with inherited mental illness, a single cycle of the procedure permitted enriched cDNAs to be visualized directly by agarose gel electrophoresis. Hybridization screening of a library of clones derived from the enriched cDNAs, employing the genomic resource as a probe, led to the identification of six novel gene fragments. This general approach to the isolation of regionally encoded genes could be applied to any subchromosomal interval as a first step towards global transcription map construction.
Brookes
A J
AJ
Molecular Genetics Section, MRC Human Genetics Unit, Western General Hospital, Edinburgh, UK.
Slorach
E M
EM
Evans
K L
KL
Thomson
M L
ML
Gosden
C M
CM
Muir
W J
WJ
Porteous
D J
DJ
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D001921:Brain / Q000378:metabolism*; D002144:Callithrix; D002415:Cats; D002417:Cattle; D002878:Chromosomes, Human, Pair 1; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs; D004587:Electrophoresis, Agar Gel; D015723:Gene Library; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011618:Psychotic Disorders / Q000235:genetics*; D011817:Rabbits; D051381:Rats; D012756:Sheep; D013045:Species Specificity; D013552:Swine; D014178:Translocation, Genetic
D018076:DNA, Complementary