Detection of Growth Hormone Gene Defects by Dideoxy Fingerprinting (ddF).

Miyata, Ichiro; Cogan, Joy D.; Prince, Melissa; Kamijo, Takashi; Ogawa, Masamichi; Phillips, John A.

doi:10.1507/endocrj.44.149

Cited by 23 publications

(21 citation statements)

References 16 publications

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“…The authenticity of the lesions detected was confirmed by repeat PCR/direct sequencing but family studies were not feasible. The novel mutations comprised two single base-pair substitutions and a gene conversion in Previously reported by Miyata et al [1997]. the promoter region, a single base-pair substitution in the 5 0 UTR, and three missense substitutions.…”

Section: Mutational Spectra In Individuals With Short Stature and Conmentioning

confidence: 93%

Novel mutations of the growth hormone 1 (GH1) gene disclosed by modulation of the clinical selection criteria for individuals with short stature

et al. 2003

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Communicated by Mark H. PaalmanSubtle mutations in the growth hormone 1 (GH1) gene have been regarded as a comparatively rare cause of short stature. Such lesions were sought in a group of 41 individuals selected for short stature, reduced height velocity, and bone age delay; a group of 11 individuals with short stature and idiopathic growth hormone deficiency (IGHD); and a group of 154 controls. Heterozygous mutations were identified in all three groups but disproportionately in the individuals with short stature, both with (odds ratio 25.2; 95% CI, 5.1-132.2) and without (odds ratio 3.6; 95% CI, 1.0-12.9) IGHD. Twenty-four novel GH1 gene lesions were found. Thirteen novel missense mutations were characterized by assaying the signal transduction activity of in vitro expressed variants; six (T27I, K41R, N47D, S71F, S108R, and T175A) exhibited a reduced ability to activate the JAK/STAT pathway. Molecular modeling suggested that both K41R and T175A might compromise GH receptor binding. Seven GH variants (R16C, K41R, S71F, E74K, Q91L, S108C, and a functional polymorphism, V110I) manifested reduced secretion in rat pituitary cells after allowance had been made for the level of expression attributable to the associated GH1 proximal promoter haplotype. A further leader peptide variant (L-11P) was not secreted. Eleven novel mutations in the GH1 gene promoter were assessed by reporter gene assay but only two, including a GH2 gene-templated gene conversion, were found to be associated with a significantly reduced level of expression. Finally, a novel intron 2 acceptor splice-site mutation, detected in a family with autosomal dominant type II IGHD, was shown to lead to the skipping of exon 3 from the GH1 transcript. A total of 15 novel GH1 gene mutations were thus considered to be of probable phenotypic significance. Such lesions are more prevalent than previously recognized and although most may be insufficient on their own to account for the observed clinical phenotype, they are nevertheless likely to play a contributory role in the etiology of short stature. Hum Mutat 21:424-440,

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Section: Mutational Spectra In Individuals With Short Stature and Conmentioning

confidence: 93%

Novel mutations of the growth hormone 1 (GH1) gene disclosed by modulation of the clinical selection criteria for individuals with short stature

et al. 2003

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“…ddF oers several advantages over SSCP, including increased sensitivity, ability to analyse larger fragments, estimation of the location of the mutation and easier, more rapid application (Martincic and Whitlock, 1996). ddF has been used to identify mutations in genes coding for the dopamine D1 receptor (Quiang et al, 1995), growth hormone (Miyata et al, 1997), monoamine oxidase (Sobell et al, 1997), x-linked severe combined immunode®ciency , BRCA1 (Durocher et al, 1996), p53 (Martincic and Whitlock, 1996;Blaszyk et al, 1995), and factor XI (Martincic et al, 1998). These studies identi®ed mutations in fragments ampli®ed from genomic DNA.…”

Section: Introductionmentioning

confidence: 99%

Detection of mutations by automated fluorescence/RNA-based dideoxy fingerprinting (ARddF)

Martincic

Koury

Gale³

et al. 1999

Oncogene

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Dideoxy ®ngerprinting (ddF) is a hybrid technique which combines aspects of single strand conformational polymorphism (SSCP) and dideoxy sequencing to detect the presence of single base changes in a de®ned fragment of nucleic acid. ddF is no more technically demanding than SSCP, yet it is more sensitive in detecting point mutations. We describe here the adaptation of conventional ddF to an automated sequencing system usinḡ uorescent Cy5 labeled primers. We show that automated RNA-based ddF (ARddF) has several advantages over conventional radioisotope-based ddF, including: (1) analysis of larger nucleic acid fragments (up to 10 3 bp), due to the ability to continuously analyse and compile sequencing information; (2) greater reliability for distinguishing mutant sequences from wild type sequences (particularly when the mutation leads to gain or loss of a dideoxy termination segment); (3) the use of¯uorescent labeled primers, making ARddF less hazardous than methods requiring radionucleotides. The use of ARddF in conjunction with new methods for isolating RNA from a small number of cells facilitates mutational analysis of small tissue biopsies and other limited samples, and will allow more widespread application of mutational screening in the setting of clinical diagnostic laboratories.

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“…Dideoxy fingerprinting (ddF) was performed as previously described [7]. We used 8 primers to complete ddF screening of the GH-1 gene (Fig.…”

Section: Methodsmentioning

confidence: 99%

Screening for Mutations in the GH-1 Gene by Dideoxy Fingerprinting (ddF)

et al. 1999

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Detection of Growth Hormone Gene Defects by Dideoxy Fingerprinting (ddF).

Cited by 23 publications

References 16 publications

Novel mutations of the growth hormone 1 (GH1) gene disclosed by modulation of the clinical selection criteria for individuals with short stature

Novel mutations of the growth hormone 1 (GH1) gene disclosed by modulation of the clinical selection criteria for individuals with short stature

Detection of mutations by automated fluorescence/RNA-based dideoxy fingerprinting (ARddF)

Screening for Mutations in the GH-1 Gene by Dideoxy Fingerprinting (ddF)

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