A new polymorphic locus, D7S411, isolated by cloning from preparative pulse-field gels is close to the mutation causing cystic fibrosis

M. Ramsay, B. J. Wainwright, M. Farrall, Xavier P. Estivill, H. Sutherland, M. F. Ho, R. Davies, S. Halford, F. Tata, C. Wicking, N. Lench, I. Bauer, C. Ferec, P. Farndon, H. Kruyer, P. Stanier, R. Williamson, P. J. Scambler

Research output: Contribution to journalArticle

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Abstract

The mutation causing cystic fibrosis (CF) has been localized to the DNA sequence of 700 kb bounded by the loci identified by the markers pMP6d-9 (D7S399) and pJ3.11 (D7S8). A 560-kb fragment obtained after SacII digestion of DNA from a cell line containing this region of human chromosome 7 in a mouse background was separated using pulse-field gel electrophoresis and isolated from the gel. The DNA was digested with BamHI prior to cloning into λ EMBL3. Approximately 0.1% of the resulting clones contained human repetitive sequences, and 24 such recombinants were studied. Of these, 23 are on chromosome 7; 8 clones were duplicated, and of the 15 different recombinants, 7 are between MET and INT1L1, and a further 7 are between INT1L1 and pMP6d-9, leaving a single marker, pG2, which is between pMP6d-9 and pJ3.11. pG2 recognizes an RFLP with XbaI. A cosmid walk from pG2 has generated a further marker, H80, which recognizes an RFLP with PstI. This new locus (D7S411) divides the remaining region between the CF flanking markers, thereby making it more accessible to fine pulse-field mapping and allowing the precise localization of further clones to this region. Although it is not possible to position the CF locus unequivocally with respect to D7S411, both polymorphic markers at this locus exhibit low but significant linkage disequilibrium with CF, placing the emphasis for the search for the gene on the D7S399 to D7S411 interval of 250 kb. None of the cystic fibrosis families that previously showed recombination with either pJ3.11 or INT1L1 is informative at the D7S411 locus.

Original languageEnglish
Pages (from-to)39-47
Number of pages9
JournalGenomics
Volume6
Issue number1
DOIs
Publication statusPublished - 1990
Externally publishedYes

Fingerprint

Cystic Fibrosis
Organism Cloning
Gels
Mutation
Chromosomes, Human, Pair 7
Clone Cells
Restriction Fragment Length Polymorphisms
Chromosomes, Human, Pair 8
Cosmids
Nucleic Acid Repetitive Sequences
DNA
Linkage Disequilibrium
Human Chromosomes
Genetic Recombination
Electrophoresis
Digestion
Cell Line
Genes

ASJC Scopus subject areas

  • Genetics

Cite this

A new polymorphic locus, D7S411, isolated by cloning from preparative pulse-field gels is close to the mutation causing cystic fibrosis. / Ramsay, M.; Wainwright, B. J.; Farrall, M.; Estivill, Xavier P.; Sutherland, H.; Ho, M. F.; Davies, R.; Halford, S.; Tata, F.; Wicking, C.; Lench, N.; Bauer, I.; Ferec, C.; Farndon, P.; Kruyer, H.; Stanier, P.; Williamson, R.; Scambler, P. J.

In: Genomics, Vol. 6, No. 1, 1990, p. 39-47.

Research output: Contribution to journalArticle

Ramsay, M, Wainwright, BJ, Farrall, M, Estivill, XP, Sutherland, H, Ho, MF, Davies, R, Halford, S, Tata, F, Wicking, C, Lench, N, Bauer, I, Ferec, C, Farndon, P, Kruyer, H, Stanier, P, Williamson, R & Scambler, PJ 1990, 'A new polymorphic locus, D7S411, isolated by cloning from preparative pulse-field gels is close to the mutation causing cystic fibrosis', Genomics, vol. 6, no. 1, pp. 39-47. https://doi.org/10.1016/0888-7543(90)90446-2
Ramsay, M. ; Wainwright, B. J. ; Farrall, M. ; Estivill, Xavier P. ; Sutherland, H. ; Ho, M. F. ; Davies, R. ; Halford, S. ; Tata, F. ; Wicking, C. ; Lench, N. ; Bauer, I. ; Ferec, C. ; Farndon, P. ; Kruyer, H. ; Stanier, P. ; Williamson, R. ; Scambler, P. J. / A new polymorphic locus, D7S411, isolated by cloning from preparative pulse-field gels is close to the mutation causing cystic fibrosis. In: Genomics. 1990 ; Vol. 6, No. 1. pp. 39-47.
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title = "A new polymorphic locus, D7S411, isolated by cloning from preparative pulse-field gels is close to the mutation causing cystic fibrosis",
abstract = "The mutation causing cystic fibrosis (CF) has been localized to the DNA sequence of 700 kb bounded by the loci identified by the markers pMP6d-9 (D7S399) and pJ3.11 (D7S8). A 560-kb fragment obtained after SacII digestion of DNA from a cell line containing this region of human chromosome 7 in a mouse background was separated using pulse-field gel electrophoresis and isolated from the gel. The DNA was digested with BamHI prior to cloning into λ EMBL3. Approximately 0.1{\%} of the resulting clones contained human repetitive sequences, and 24 such recombinants were studied. Of these, 23 are on chromosome 7; 8 clones were duplicated, and of the 15 different recombinants, 7 are between MET and INT1L1, and a further 7 are between INT1L1 and pMP6d-9, leaving a single marker, pG2, which is between pMP6d-9 and pJ3.11. pG2 recognizes an RFLP with XbaI. A cosmid walk from pG2 has generated a further marker, H80, which recognizes an RFLP with PstI. This new locus (D7S411) divides the remaining region between the CF flanking markers, thereby making it more accessible to fine pulse-field mapping and allowing the precise localization of further clones to this region. Although it is not possible to position the CF locus unequivocally with respect to D7S411, both polymorphic markers at this locus exhibit low but significant linkage disequilibrium with CF, placing the emphasis for the search for the gene on the D7S399 to D7S411 interval of 250 kb. None of the cystic fibrosis families that previously showed recombination with either pJ3.11 or INT1L1 is informative at the D7S411 locus.",
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T1 - A new polymorphic locus, D7S411, isolated by cloning from preparative pulse-field gels is close to the mutation causing cystic fibrosis

AU - Ramsay, M.

AU - Wainwright, B. J.

AU - Farrall, M.

AU - Estivill, Xavier P.

AU - Sutherland, H.

AU - Ho, M. F.

AU - Davies, R.

AU - Halford, S.

AU - Tata, F.

AU - Wicking, C.

AU - Lench, N.

AU - Bauer, I.

AU - Ferec, C.

AU - Farndon, P.

AU - Kruyer, H.

AU - Stanier, P.

AU - Williamson, R.

AU - Scambler, P. J.

PY - 1990

Y1 - 1990

N2 - The mutation causing cystic fibrosis (CF) has been localized to the DNA sequence of 700 kb bounded by the loci identified by the markers pMP6d-9 (D7S399) and pJ3.11 (D7S8). A 560-kb fragment obtained after SacII digestion of DNA from a cell line containing this region of human chromosome 7 in a mouse background was separated using pulse-field gel electrophoresis and isolated from the gel. The DNA was digested with BamHI prior to cloning into λ EMBL3. Approximately 0.1% of the resulting clones contained human repetitive sequences, and 24 such recombinants were studied. Of these, 23 are on chromosome 7; 8 clones were duplicated, and of the 15 different recombinants, 7 are between MET and INT1L1, and a further 7 are between INT1L1 and pMP6d-9, leaving a single marker, pG2, which is between pMP6d-9 and pJ3.11. pG2 recognizes an RFLP with XbaI. A cosmid walk from pG2 has generated a further marker, H80, which recognizes an RFLP with PstI. This new locus (D7S411) divides the remaining region between the CF flanking markers, thereby making it more accessible to fine pulse-field mapping and allowing the precise localization of further clones to this region. Although it is not possible to position the CF locus unequivocally with respect to D7S411, both polymorphic markers at this locus exhibit low but significant linkage disequilibrium with CF, placing the emphasis for the search for the gene on the D7S399 to D7S411 interval of 250 kb. None of the cystic fibrosis families that previously showed recombination with either pJ3.11 or INT1L1 is informative at the D7S411 locus.

AB - The mutation causing cystic fibrosis (CF) has been localized to the DNA sequence of 700 kb bounded by the loci identified by the markers pMP6d-9 (D7S399) and pJ3.11 (D7S8). A 560-kb fragment obtained after SacII digestion of DNA from a cell line containing this region of human chromosome 7 in a mouse background was separated using pulse-field gel electrophoresis and isolated from the gel. The DNA was digested with BamHI prior to cloning into λ EMBL3. Approximately 0.1% of the resulting clones contained human repetitive sequences, and 24 such recombinants were studied. Of these, 23 are on chromosome 7; 8 clones were duplicated, and of the 15 different recombinants, 7 are between MET and INT1L1, and a further 7 are between INT1L1 and pMP6d-9, leaving a single marker, pG2, which is between pMP6d-9 and pJ3.11. pG2 recognizes an RFLP with XbaI. A cosmid walk from pG2 has generated a further marker, H80, which recognizes an RFLP with PstI. This new locus (D7S411) divides the remaining region between the CF flanking markers, thereby making it more accessible to fine pulse-field mapping and allowing the precise localization of further clones to this region. Although it is not possible to position the CF locus unequivocally with respect to D7S411, both polymorphic markers at this locus exhibit low but significant linkage disequilibrium with CF, placing the emphasis for the search for the gene on the D7S399 to D7S411 interval of 250 kb. None of the cystic fibrosis families that previously showed recombination with either pJ3.11 or INT1L1 is informative at the D7S411 locus.

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