Mapping of the human nicotinic acetylcholine receptor [beta]3 gene (CHRNB3) within chromosome 8p11. 2
- Cancer Institute, Tokyo (Japan)
The authors have used an exon amplification method to construct a transcriptional map for human chromosome 8. With this method, transcribed sequences from defined regions of genomic DNA can be efficiently isolated using cosmid clones mapped to human chromosome 8. Cosmid DNAs were digested with BglII and BamHI and ligated into a BamHI site of an exon trapping vector, pSPL1. Transfection of the subcloned DNAs into Cos7 cells, isolation of cytoplasmic RNA, synthesis of cDNA by reverse transcriptase, and amplification of spliced fragments were performed according to the method described by Buckler et al. Amplified fragments were subcloned into a plasmid, pBluescriptII, and sequenced by the dideoxy chain termination method. Sequence analysis to search for similarity or identity to known genes with the program FASTA detected complete identity of one (ET634-2) of these exon amplification fragments, 227 bp in length, to the nucleotide sequence at 1138-1364 of the cDNA for the human nicotinic acetylcholine receptor (nAChR) [beta]3 gene. This transcribed fragment, containing a part of the human nAChR [beta]3 gene, was isolated from cosmid clone cCI8-328, which was previously mapped to 8p11.2 by fluorescence in situ hybridization. Localization of this gene to chromosome agreed with the results of previous mapping experiments using somatic hybrid cell lines.
- OSTI ID:
- 7101440
- Journal Information:
- Genomics; (United States), Journal Name: Genomics; (United States) Vol. 21:2; ISSN 0888-7543; ISSN GNMCEP
- Country of Publication:
- United States
- Language:
- English
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