A recombination-based assay demonstrates that the fragile X sequence is transcribed widely during development.
To identify transcribed sequences rapidly and efficiently, we have developed a recombination-based assay to screen bacteriophage lambda libraries for sequences that share homology with a given probe. This strategy determines analytically whether a given probe is transcribed in a given tissue at a given time of development, and may also be used to isolate preparatively the transcribed sequence free of the screening probe. We illustrate this technology for the fragile X sequence, demonstrating that it is transcribed ubiquitously in an 11 week fetus, in a variety of 20 week human fetal tissues, including brain, spinal cord, eye, liver, kidney and skeletal muscle, and in adult jejunum.
Hanzlik
A J
AJ
University of Michigan Medical School, Howard Hughes Medical Institute, Ann Arbor 48109-0650.
Osemlak-Hanzlik
M M
MM
Hauser
M A
MA
Kurnit
D M
DM
D000328:Adult; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D004247:DNA; D015342:DNA Probes; D005333:Fetus / Q000378:metabolism*; D051860:Fragile X Messenger Ribonucleoprotein 1; D005600:Fragile X Syndrome / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D005821:Genetic Techniques; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000235:genetics; D009928:Organ Specificity / Q000235:genetics; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins; D011995:Recombination, Genetic; D014158:Transcription, Genetic
D004247:DNA; D015342:DNA Probes; D051860:Fragile X Messenger Ribonucleoprotein 1; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; C497683:FMR1 protein, human