skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Marker development for the EPM1 region of human chromosome 21, q22.3

Abstract

New STSs have been developed for a 0.9 Mb region of chromosome 21 that is not represented in existing YAC libraries using an efficient method that is generally applicable to any region of the genome. The region, 21q22.3, is of particular interest because the gene for progressive myoclonic epilepsy of the Unverricht-Lundborg type (EPM1) maps to this region. Until recently there were only three probes for the 1.3 Mb surrounding the EPM1 gene (D21S141,LJ112, LB2T). This very limited number of probes is problematic for obtaining clone coverage and for confirming map position of newly developed markers in the EPM1 region. To develop new markers, a somatic cell hybrid containing chromosome 21 as its only human complement (GMO8854) was digested with NOT1 and hybridized with D21S141. The fragment hybridizing with D21S141 was excised, amplified by Alu-PCR and the amplification products were cloned and sequenced. Of the fifteen clones sequenced, four were duplicates and one consisted entirely of repeat sequences. STSs were developed for the remaining ten unique clones. To determine the map position of the new STSs, quantitive PCR was used in conjunction with whole genome radiation hybrid (RH) mapping. Quantitative PCR confirmed that the STSs mapped to appropriately sized PFGEmore » fragments and whole genome RH mapping showed that the makers were linked and gave order and distance information. Three of the new STSs are in the EPM1 region, providing additional starting points for obtaining clone coverage and gene isolation. This combination of techniques for developing markers and confirming map position is an effective approach for obtaining probes and has general applicability for regions of the genome not represented in YAC or cosmid libraries.« less

Authors:
; ;  [1]
  1. Stanford Univ., CA (United States); and others
Publication Date:
OSTI Identifier:
134492
Report Number(s):
CONF-941009-
Journal ID: AJHGAG; ISSN 0002-9297; TRN: 95:005313-1226
Resource Type:
Journal Article
Journal Name:
American Journal of Human Genetics
Additional Journal Information:
Journal Volume: 55; Journal Issue: Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; HUMAN CHROMOSOME 21; GENETIC MAPPING; DNA SEQUENCING; DNA-CLONING; EFFICIENCY; GENES; EPILEPSY; PATIENTS; HEREDITARY DISEASES; PROBES; DESIGN; SOMATIC CELLS; HYBRIDIZATION; POLYMERASE CHAIN REACTION; BIOLOGICAL MARKERS; YEASTS; DNA HYBRIDIZATION; ELECTROPHORESIS

Citation Formats

Warrington, I A, O`Connor, K, and Hebert, S. Marker development for the EPM1 region of human chromosome 21, q22.3. United States: N. p., 1994. Web.
Warrington, I A, O`Connor, K, & Hebert, S. Marker development for the EPM1 region of human chromosome 21, q22.3. United States.
Warrington, I A, O`Connor, K, and Hebert, S. 1994. "Marker development for the EPM1 region of human chromosome 21, q22.3". United States.
@article{osti_134492,
title = {Marker development for the EPM1 region of human chromosome 21, q22.3},
author = {Warrington, I A and O`Connor, K and Hebert, S},
abstractNote = {New STSs have been developed for a 0.9 Mb region of chromosome 21 that is not represented in existing YAC libraries using an efficient method that is generally applicable to any region of the genome. The region, 21q22.3, is of particular interest because the gene for progressive myoclonic epilepsy of the Unverricht-Lundborg type (EPM1) maps to this region. Until recently there were only three probes for the 1.3 Mb surrounding the EPM1 gene (D21S141,LJ112, LB2T). This very limited number of probes is problematic for obtaining clone coverage and for confirming map position of newly developed markers in the EPM1 region. To develop new markers, a somatic cell hybrid containing chromosome 21 as its only human complement (GMO8854) was digested with NOT1 and hybridized with D21S141. The fragment hybridizing with D21S141 was excised, amplified by Alu-PCR and the amplification products were cloned and sequenced. Of the fifteen clones sequenced, four were duplicates and one consisted entirely of repeat sequences. STSs were developed for the remaining ten unique clones. To determine the map position of the new STSs, quantitive PCR was used in conjunction with whole genome radiation hybrid (RH) mapping. Quantitative PCR confirmed that the STSs mapped to appropriately sized PFGE fragments and whole genome RH mapping showed that the makers were linked and gave order and distance information. Three of the new STSs are in the EPM1 region, providing additional starting points for obtaining clone coverage and gene isolation. This combination of techniques for developing markers and confirming map position is an effective approach for obtaining probes and has general applicability for regions of the genome not represented in YAC or cosmid libraries.},
doi = {},
url = {https://www.osti.gov/biblio/134492}, journal = {American Journal of Human Genetics},
number = Suppl.3,
volume = 55,
place = {United States},
year = {Thu Sep 01 00:00:00 EDT 1994},
month = {Thu Sep 01 00:00:00 EDT 1994}
}