Detection of a novel deletion in the cystathionine beta-synthase (CBS) gene using an improved genomic DNA based method

M Gaustadnes; L A Kluijtmans; O K Jensen; K Rasmussen; S G Heil; J P Kraus; H J Blom; J Ingerslev; N Rüdiger; Ole Kudsk Jensen

Detection of a novel deletion in the cystathionine beta-synthase (CBS) gene using an improved genomic DNA based method

M Gaustadnes, L A Kluijtmans, O K Jensen, K Rasmussen, S G Heil, J P Kraus, H J Blom, J Ingerslev, N Rüdiger, Ole Kudsk Jensen

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

14 Citations (Scopus)

Abstract

We elucidated the intron-exon boundaries of the 15 coding exons of the human cystathionine beta-synthase (CBS) gene in order to establish an improved method based on PCR and direct sequencing for detection of CBS mutations. Using this method we identified the pathogenic mutations in two Danish siblings with CBS deficiency. Patients were compound heterozygotes: we detected the 833T-->C mutation and a novel 22 bp deletion of exon 4 (493-514del) that introduces a frameshift and a stop codon immediately after the deletion. The deletion resulted in no detectable mRNA from this allele, as assessed by sequencing of cDNA. The established method represents an improvement of the existing method based on sequencing of cDNA because it permits the detection of mutations within the entire coding region of the CBS gene from a peripheral blood sample, including splice mutations and mutations resulting in the lack or a reduced amount of transcript.

Original language	English
Journal	FEBS Letters
Volume	431
Issue	2
Pages (from-to)	175-9
Number of pages	5
ISSN	0014-5793
Publication status	Published - 17 Jul 1998

Keywords

Adult
Base Sequence
Cystathionine beta-Synthase
DNA Mutational Analysis
DNA Primers
Exons
Female
Fibroblasts
Gene Deletion
Genetic Testing
Homocystinuria
Humans
Introns
Male
Molecular Sequence Data
Pedigree
Polymerase Chain Reaction

Cite this

@article{2872c5eae3b44dfc9d0d38ef91edda7b,

title = "Detection of a novel deletion in the cystathionine beta-synthase (CBS) gene using an improved genomic DNA based method",

abstract = "We elucidated the intron-exon boundaries of the 15 coding exons of the human cystathionine beta-synthase (CBS) gene in order to establish an improved method based on PCR and direct sequencing for detection of CBS mutations. Using this method we identified the pathogenic mutations in two Danish siblings with CBS deficiency. Patients were compound heterozygotes: we detected the 833T-->C mutation and a novel 22 bp deletion of exon 4 (493-514del) that introduces a frameshift and a stop codon immediately after the deletion. The deletion resulted in no detectable mRNA from this allele, as assessed by sequencing of cDNA. The established method represents an improvement of the existing method based on sequencing of cDNA because it permits the detection of mutations within the entire coding region of the CBS gene from a peripheral blood sample, including splice mutations and mutations resulting in the lack or a reduced amount of transcript.",

keywords = "Adult, Base Sequence, Cystathionine beta-Synthase, DNA Mutational Analysis, DNA Primers, Exons, Female, Fibroblasts, Gene Deletion, Genetic Testing, Homocystinuria, Humans, Introns, Male, Molecular Sequence Data, Pedigree, Polymerase Chain Reaction",

author = "M Gaustadnes and Kluijtmans, {L A} and Jensen, {O K} and K Rasmussen and Heil, {S G} and Kraus, {J P} and Blom, {H J} and J Ingerslev and N R{\"u}diger and Jensen, {Ole Kudsk}",

year = "1998",

month = jul,

day = "17",

language = "English",

volume = "431",

pages = "175--9",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "John Wiley & Sons Inc.",

number = "2",

}

Detection of a novel deletion in the cystathionine beta-synthase (CBS) gene using an improved genomic DNA based method. / Gaustadnes, M; Kluijtmans, L A; Jensen, O K et al.
In: FEBS Letters, Vol. 431, No. 2, 17.07.1998, p. 175-9.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Detection of a novel deletion in the cystathionine beta-synthase (CBS) gene using an improved genomic DNA based method

AU - Gaustadnes, M

AU - Kluijtmans, L A

AU - Jensen, O K

AU - Rasmussen, K

AU - Heil, S G

AU - Kraus, J P

AU - Blom, H J

AU - Ingerslev, J

AU - Rüdiger, N

AU - Jensen, Ole Kudsk

PY - 1998/7/17

Y1 - 1998/7/17

N2 - We elucidated the intron-exon boundaries of the 15 coding exons of the human cystathionine beta-synthase (CBS) gene in order to establish an improved method based on PCR and direct sequencing for detection of CBS mutations. Using this method we identified the pathogenic mutations in two Danish siblings with CBS deficiency. Patients were compound heterozygotes: we detected the 833T-->C mutation and a novel 22 bp deletion of exon 4 (493-514del) that introduces a frameshift and a stop codon immediately after the deletion. The deletion resulted in no detectable mRNA from this allele, as assessed by sequencing of cDNA. The established method represents an improvement of the existing method based on sequencing of cDNA because it permits the detection of mutations within the entire coding region of the CBS gene from a peripheral blood sample, including splice mutations and mutations resulting in the lack or a reduced amount of transcript.

AB - We elucidated the intron-exon boundaries of the 15 coding exons of the human cystathionine beta-synthase (CBS) gene in order to establish an improved method based on PCR and direct sequencing for detection of CBS mutations. Using this method we identified the pathogenic mutations in two Danish siblings with CBS deficiency. Patients were compound heterozygotes: we detected the 833T-->C mutation and a novel 22 bp deletion of exon 4 (493-514del) that introduces a frameshift and a stop codon immediately after the deletion. The deletion resulted in no detectable mRNA from this allele, as assessed by sequencing of cDNA. The established method represents an improvement of the existing method based on sequencing of cDNA because it permits the detection of mutations within the entire coding region of the CBS gene from a peripheral blood sample, including splice mutations and mutations resulting in the lack or a reduced amount of transcript.

KW - Adult

KW - Base Sequence

KW - Cystathionine beta-Synthase

KW - DNA Mutational Analysis

KW - DNA Primers

KW - Exons

KW - Female

KW - Fibroblasts

KW - Gene Deletion

KW - Genetic Testing

KW - Homocystinuria

KW - Humans

KW - Introns

KW - Male

KW - Molecular Sequence Data

KW - Pedigree

KW - Polymerase Chain Reaction

M3 - Journal article

C2 - 9708897

SN - 0014-5793

VL - 431

SP - 175

EP - 179

JO - FEBS Letters

JF - FEBS Letters

IS - 2

ER -

Detection of a novel deletion in the cystathionine beta-synthase (CBS) gene using an improved genomic DNA based method

Abstract

Keywords

Fingerprint

Cite this