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Title: Identification of neurofibromatosis 1 (NF1) homologous loci by direct sequencing, fluorescence in situ hybridization, and PCR amplification of somatic cell hybrids

Abstract

Using fluorescence in situ hybridization (FISH), we have identified seven NF1-related loci, two separate loci on chromosome 2, at bands 2q21 and 2q33-q34, and one locus each on five other chromosomes at bands 14q11.2, 15q11.2, 18p11.2, 21q11.2-q21, and 22q11.2. Application of PCR using NF1 primer pairs and genomic DNA from somatic cell hybrids confirmed the above loci, identified additional loci on chromosomes 12 and 15, and showed that the various loci do not share homology beyond NF1 exon 27b. Sequenced PCR products representing segments corresponding to NF1 exons from these loci demonstrated greater than 95% sequence identity with the NF1 locus. We used sequence differences between bona fide NF1 and NF1-homologous loci to strategically design primer sets to specifically amplify 30 of 36 exons within the 5{prime} end of the NF1 gene. These developments have facilitated mutation analysis at the NF1 locus using genomic DNA as template. 41 refs., 3 figs., 3 tabs.

Authors:
; ;  [1]
  1. Univ. of Utah, Salt Lake City (United States)
Publication Date:
OSTI Identifier:
437166
Resource Type:
Journal Article
Journal Name:
Genomics
Additional Journal Information:
Journal Volume: 30; Journal Issue: 3; Other Information: PBD: 10 Dec 1995
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; GENES; GENETIC MAPPING; DNA SEQUENCING; GENE MUTATIONS; PATIENTS; HEREDITARY DISEASES; NERVOUS SYSTEM DISEASES; HUMAN CHROMOSOME 2; HUMAN CHROMOSOME 14; HUMAN CHROMOSOME 15; HUMAN CHROMOSOME 18; HUMAN CHROMOSOME 21; HUMAN CHROMOSOME 22; FLUORESCENCE; DNA HYBRIDIZATION; SOMATIC CELLS; POLYMERASE CHAIN REACTION

Citation Formats

Purandare, S M, Neil, S M, Brothman, A, and Jikei Univ., Tokyo. Identification of neurofibromatosis 1 (NF1) homologous loci by direct sequencing, fluorescence in situ hybridization, and PCR amplification of somatic cell hybrids. United States: N. p., 1995. Web. doi:10.1006/geno.1995.1268.
Purandare, S M, Neil, S M, Brothman, A, & Jikei Univ., Tokyo. Identification of neurofibromatosis 1 (NF1) homologous loci by direct sequencing, fluorescence in situ hybridization, and PCR amplification of somatic cell hybrids. United States. https://doi.org/10.1006/geno.1995.1268
Purandare, S M, Neil, S M, Brothman, A, and Jikei Univ., Tokyo. 1995. "Identification of neurofibromatosis 1 (NF1) homologous loci by direct sequencing, fluorescence in situ hybridization, and PCR amplification of somatic cell hybrids". United States. https://doi.org/10.1006/geno.1995.1268.
@article{osti_437166,
title = {Identification of neurofibromatosis 1 (NF1) homologous loci by direct sequencing, fluorescence in situ hybridization, and PCR amplification of somatic cell hybrids},
author = {Purandare, S M and Neil, S M and Brothman, A and Jikei Univ., Tokyo},
abstractNote = {Using fluorescence in situ hybridization (FISH), we have identified seven NF1-related loci, two separate loci on chromosome 2, at bands 2q21 and 2q33-q34, and one locus each on five other chromosomes at bands 14q11.2, 15q11.2, 18p11.2, 21q11.2-q21, and 22q11.2. Application of PCR using NF1 primer pairs and genomic DNA from somatic cell hybrids confirmed the above loci, identified additional loci on chromosomes 12 and 15, and showed that the various loci do not share homology beyond NF1 exon 27b. Sequenced PCR products representing segments corresponding to NF1 exons from these loci demonstrated greater than 95% sequence identity with the NF1 locus. We used sequence differences between bona fide NF1 and NF1-homologous loci to strategically design primer sets to specifically amplify 30 of 36 exons within the 5{prime} end of the NF1 gene. These developments have facilitated mutation analysis at the NF1 locus using genomic DNA as template. 41 refs., 3 figs., 3 tabs.},
doi = {10.1006/geno.1995.1268},
url = {https://www.osti.gov/biblio/437166}, journal = {Genomics},
number = 3,
volume = 30,
place = {United States},
year = {Sun Dec 10 00:00:00 EST 1995},
month = {Sun Dec 10 00:00:00 EST 1995}
}