Identification of sequences of chromosome 7 that are expressed in sweat gland epithelial cells.
This paper describes an approach that can be used to identify specifically expressed coding sequences in defined regions of genomic DNA. We developed this method to identify expressed sequences from chromosome 7 located at or near the cystic fibrosis (CF) locus. Radioactively labelled single-stranded cDNAs derived from sweat gland epithelial cells and from fibroblasts were used to screen a genomic library constructed from flow-sorted chromosomes. Differential screening of phage lifts with these two probes yielded 36 different DNA segments. By using somatic cell hybrids containing different portions of chromosome 7, four of the clones were mapped to the 7q31 region in which the CF locus is located. These four clones and two others that gave strong differential epithelial signals but that were not within 7q31 were studied further. Restriction fragment length polymorphisms (RFLPs) were identified for two of the DNA segments within 7q31 and used for linkage analysis using a panel of CF families. One DNA segment was assigned to a location centromeric to the met locus. The other marker did not show recombination with CF but was subsequently excluded from the CF region by physical mapping. Three of the six DNA segments were found to hybridize to various RNAs using the Northern technique and therefore contain portions of genes. One of the clones showed strong differential expression when epithelial tissues were compared to fibroblasts and may represent an epithelium-specific gene.
Burns
J
J
Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
Melmer
G
G
Rommens
J M
JM
Riordan
J R
JR
Buchwald
M
M
D001345:Autoradiography; D015152:Blotting, Northern; D002874:Chromosome Mapping; D002897:Chromosomes, Human, Pair 7 / Q000032:analysis*; D003550:Cystic Fibrosis / Q000235:genetics*; D015342:DNA Probes; D004848:Epithelium / Q000032:analysis; D008040:Genetic Linkage; D015894:Genome, Human; D006801:Humans; D009693:Nucleic Acid Hybridization; D012150:Polymorphism, Restriction Fragment Length; D013545:Sweat Glands / Q000032:analysis*
D015342:DNA Probes