Novel Mutation of the Initiation Codon of PAX9 Causes Oligodontia

2005 ◽  
Vol 84 (1) ◽  
pp. 43-47 ◽  
Author(s):  
M.L. Klein ◽  
P. Nieminen ◽  
L. Lammi ◽  
E. Niebuhr ◽  
S. Kreiborg
Keyword(s):  
Tooth Development ◽  
Novel Mutation ◽  
Direct Sequencing ◽  
Single Gene ◽  
Third Molar ◽  
Initiation Codon ◽  
Permanent Teeth ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Exon 1

Tooth development is under strict genetic control. Oligodontia is defined as the congenital absence of 6 or more permanent teeth, excluding the third molar. The occurrence of non-syndromic oligodontia is poorly understood, but in recent years several cases have been described where a single gene mutation is associated with oligodontia. Several studies have shown that MSX1 and PAX9 play a role in early tooth development. We screened one family with non-syndromic oligodontia for mutations in MSX1 and PAX9. The pedigree showed an autosomal-dominant pattern of inheritance. Direct sequencing and restriction enzyme analysis revealed a novel heterozygous A to G transition mutation in the AUG initiation codon of PAX9 in exon 1 in the affected members of the family. This is the first mutation found in the initiation codon of PAX9, and we suggest that it causes haploinsufficiency.

Hereditary Protein C Deficiency Associated with Mutations in Exon IX of the Protein C Gene

1994 ◽  
Vol 72 (02) ◽  
pp. 203-208 ◽  
Author(s):  
R G Doig ◽  
C G Begley ◽  
K M McGrath
Keyword(s):  
Amino Acid ◽  
Protein C ◽  
Novel Mutation ◽  
Single Point ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Single Point Mutation ◽  
Nucleotide Position ◽  
Type I ◽  
Protein C Deficiency

SummaryThis report describes five families with symptomatic hereditary protein C deficiency. Using a polymerase chain reaction (PCR)-based method, the entire coding sequence and intron-exon boundaries of the protein C gene was amplified from genomic DNA. In each family a single point mutation in the protein C gene was identified. Two unrelated families were found to share the same mutation, while the other three had different mutations. In the first two families with type I protein C deficiency the normal cytosine residue at nucleotide position 8551 in the protein C gene was replaced by thymidine leading to substitution of the normal proline residue at amino acid position 279 by leucine. In the third family with type I deficiency a previously undescribed mutation was identified. In this family the guanosine residue at position 8559 was replaced by adenosine (glycine 282 substituted by serine). In the fourth family, also with type I deficiency, guanosine 8589 was replaced by adenosine (glycine 292 substituted by serine). The fifth family had type II deficiency and in affected members cytosine 8769 was replaced by thymidine (arginine 352 substituted by tryptophan). All these mutations lead to amino acid substitutions in the serine protease domain of the mature protein. All were able to be confirmed by restriction enzyme analysis of PCR-derived DNA. In addition the novel mutation at nucleotide position 8559 was also demonstrable using single strand conformation polymorphism (SSCP) analysis of PCR-derived DNA. These mutations were likely examples of deamination of methylated cytosine occurring in cytosine-phosphate-guanosine (CpG) dinucleotide sequences. These findings confirm the genetic heterogeneity of hereditary protein C deficiency in these families.

scholarly journals Genetic Analysis of the CYP2D6 Locus in a Hong Kong Chinese Population

2000 ◽  
Vol 46 (1) ◽  
pp. 18-23 ◽  
Author(s):  
Mercè Garcia-Barceló ◽  
Lok Yee Chow ◽  
Helen Fung Kum Chiu ◽  
Yun Kowk Wing ◽  
Dominic Tak Shing Lee ◽  
...  
Keyword(s):  
Hong Kong ◽  
Chinese Population ◽  
Hong Kong Chinese ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Chinese Populations ◽  
Specific Pcr ◽  
Significant Difference ◽  
Exon 1 ◽  
Allele Specific

Abstract Background: The cytochrome P450 CYP2D6 enzyme debrisoquine 4-hydroxylase metabolizes many different classes of commonly used drugs, such as tricyclic antidepressants and neuroleptics. Genetic polymorphism of the CYP2D6 gene is responsible for pronounced interindividual and interracial differences in the metabolism of these drugs. The CYP2D6*10 allele and its variants are the most frequent alleles found in Orientals, and they are responsible for diminished debrisoquine 4-hydroxylase activity because of the presence of a C188→T mutation in exon 1. Methods: One hundred nineteen Hong Kong Chinese subjects were genotyped by means of allele-specific PCR, PCR, and restriction enzyme analysis for 10 CYP2D6 alleles (CYP2D6*1, *2, *4D, *5, *8/*14, *10A, *10B, *15, *16, and J9). Results: CYP2D6*10B was the most prevalent allele, and CYP2D6*10/CYP2D6*10 was the most frequent genotype, representing 46.22% of the population. Conclusions: There was no significant difference in the prevalence of the alleles analyzed between our study and the Chinese populations genotyped previously. This is the largest study in terms of the number of CYP2D6 alleles analyzed in an Oriental population and the first one conducted in a Hong Kong Chinese population.

A novel mutation in the transmembrane region of glycoprotein IX associated with Bernard-Soulier syndrome

2004 ◽  
Vol 92 (09) ◽  
pp. 606-613 ◽  
Author(s):  
Xiaojuan Zhao ◽  
Weiming Duan ◽  
Jianxin Fu ◽  
Mingen Lu ◽  
Giamin Wang ◽  
...  
Keyword(s):  
Cho Cells ◽  
Novel Mutation ◽  
Flow Cytometric Analysis ◽  
Chinese Hamster ◽  
Surface Expression ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Wild Type ◽  
Transmembrane Region ◽  
Bernard Soulier Syndrome

SummaryWe describe here a novel mutation in glycoprotein (GP) IX transmembrane region in a patient with Bernard-Soulier syndrome (BSS). Flow cytometric analysis of the patient’s platelets showed that GP Iba and GP IX were expressed at decreased levels. Sequence analysis of the gene coding for GP IX revealed a homozygous (G to A) transition at nucleotide 2113, resulting in a Ala 140 (GCC) to Thr (ACC) replacement in the mature peptide, whereas no defects were found in the coding region of the GP Iba and GP Ib? gene. Allele-specific restriction enzyme analysis using HPYCH4 III revealed that the patient was homozygous and her mother and brother were heterozygous for the defect, and excluded the possibility that the mutation was a polymorphism of GP IX.To clarify the effect of this mutation on the surface expression of the GP Ib/IX complex, we introduced this mutation into the cDNA of GP IX by site-directed mutagenesis and performed in vitro transfection studies with plasmids harboring GP Iba, GP Ib? and wild-type GP IX or mutant GP IX. Mutant GP IX decreased the surface expression of GP Iba and GP IX, whereas both immunostaining and immunoblotting of the transfected Chinese hamster ovary (CHO) cells showed abundant GP Iba and GP IX in the cytoplasm of the CHO cells transfected with plasmids harboring GP Iba, GP Ib? and wild-type GP IX or mutant GP IX These findings indicate that the Ala140→Thr mutation in the transmembrane region of GP IX does not induce intracellular GP Ib/IX complex degradation, but prevents its insertion in the cytoplasmic membrane of platelets and CHO cells.

A New Candidate Missense Mutation (Leu 1657 Ile) in an Apparently Asymptomatic Type 2A (Phenotype IIA) von Willebrand Disease Family

2000 ◽  
Vol 84 (09) ◽  
pp. 369-373 ◽  
Author(s):  
A. M. Guilliatt ◽  
G. K. Surdhar ◽  
B. D. M. Theophilus ◽  
F. G. H. Hill ◽  
M. S. Enayat
Keyword(s):  
Missense Mutation ◽  
Direct Sequencing ◽  
Mutation Screening ◽  
Von Willebrand Disease ◽  
Site Directed Mutagenesis ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Full Spectrum ◽  
Von Willebrand

SummaryType 2A von Willebrand disease (VWD) is mostly an autosomal dominantly inherited bleeding disorder characterised by a qualitative defect of von Willebrand factor (VWF). Mutation screening was used to screen the whole of VWF gene followed by direct sequencing to detect the mutation in a father and son diagnosed with type 2A (phenotype IIA) von Willebrand disease. A C5219 to A transversion was detected predicting Leucine to Isoleucine substitution in codon 1657. This novel missense mutation which was also identified by MboI restriction enzyme analysis, was found in both patient and his father but not in any other unaffected family member or 50 unrelated normal individuals. This substitution was reproduced by in vitro site directed mutagenesis of full-length VWF cDNA and transiently expressed in COS-7 cells. The corresponding recombinant VWF protein exhibited the full spectrum of VWF multimers, suggesting that the abnormal multimer seen in the patient results from increased proteolysis.

scholarly journals A Novel Mutation in the Preprovasopressin Gene Identified in a Kindred with Autosomal Dominant Neurohypophyseal Diabetes Insipidus

2004 ◽  
Vol 89 (4) ◽  
pp. 1963-1968 ◽  
Author(s):  
Justin T. Wahlstrom ◽  
Michael J. Fowler ◽  
Wendell E. Nicholson ◽  
William J. Kovacs
Keyword(s):  
Diabetes Insipidus ◽  
Water Conservation ◽  
Autosomal Dominant ◽  
Novel Mutation ◽  
Single Gene ◽  
Free Water ◽  
Urinary Concentration ◽  
Gene Products ◽  
Cellular Toxicity ◽  
Exon 1

Abstract Autosomal dominant neurohypophyseal diabetes insipidus (ADNDI) is a defect in free water conservation caused by mutations in the single gene that encodes both vasopressin (VP) and its binding protein, neurophysin II (NP II). Most of the human mutations in this gene have been in the portion encoding the NP molecule; the resultant abnormal gene products are believed to cause cellular toxicity as improperly folded precursor molecules accumulate in the endoplasmic reticulum. We identified a new American kindred with ADNDI and found a novel mutation in the VP molecule. A 78-yr-old man was noted to have hypotonic polyuria and plasma hyperosmolarity; the urinary concentration defect was reversed by administration of VP. His symptomatology dated to childhood, and his family history was consistent with autosomal transmission of the polyuric syndrome, with affected members in three generations, including several females. Affected individuals were found to be heterozygous for a 3-bp deletion in exon 1 of arginine VP (AVP)-NP II, predicting a deletion of phenylalanine 3 (known to be critical for receptor binding) in the VP nonapeptide. Neuro 2A cells stably transfected with the mutant AVP-NP construct showed increased rates of apoptosis as assessed by flow cytometric methods. These observations support the concept that cellular toxicity of abnormal AVP-NP gene products underlies the development of ADNDI, and the data further demonstrate that mutations affecting the AVP moiety can result in initiation of these pathological processes.

scholarly journals A molecular characterization ofClostridium difficileisolates from humans, animals and their environments

1993 ◽  
Vol 111 (2) ◽  
pp. 257-264 ◽  
Author(s):  
G. O'Neill ◽  
J. E. Adams ◽  
R. A. Bowman ◽  
T. V. Riley
Keyword(s):  
Fragment Length Polymorphism ◽  
Hospital Environment ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Direct Transmission ◽  
Chromosomal Dna ◽  
Veterinary Clinic ◽  
Enhanced Chemiluminescence ◽  
Typing Methods ◽  
Rflp Typing

SummaryIt is generally accepted that most patients withClostridium difficile-associated diarrhoea acquire the organism from the environment. Recently we demonstrated that household pets may constitute a significant reservoir ofC. difficilethrough gastrointestinal carriage in up to 39% of cats and dogs. These findings suggested that direct transmission from household pets, or contamination of the environment by them, may be a factor in the pathogenesis ofC. difficile-associated diarrhoea. To investigate this possibility, we examined isolates ofC. difficilefrom humans, pets and the environment by restriction enzyme analysis (REA) and restriction fragment length polymorphism (RFLP) typing using enhanced chemiluminescence. Both REA and RFLP typing methods usedHindIII digests of chromosomal DNA. A total of 116 isolates ofC. difficilefrom pets (26), veterinary clinic environmental sites (33), humans (37) and hospital environmental sites (20) was examined. REA was far more discriminatory than RFLP typing and for all isolates there were 34 REA types versus 6 RFLP types. There was good correlation between the REA types found in isolates from pets and from the veterinary clinic environment, and between isolates from humans and from those found in the hospital environment. There was, however, no correlation between REA type ofC. difficilefound in pets and isolates of human origin. We conclude that there may still be a risk of humans acquiringC. difficilefrom domestic pets as these findings may be the result of geographical variation.

scholarly journals Specific Detection of Xanthomonas axonopodis pv. dieffenbachiae in Anthurium (Anthurium andreanum) Tissues by Nested PCR

2006 ◽  
Vol 72 (2) ◽  
pp. 1072-1078 ◽  
Author(s):  
Isabelle Robène-Soustrade ◽  
Philippe Laurent ◽  
Lionel Gagnevin ◽  
Emmanuel Jouen ◽  
Olivier Pruvost
Keyword(s):  
Diagnostic Tool ◽  
Nested Pcr ◽  
Restriction Analysis ◽  
Detection Threshold ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Amplification Product ◽  
Xanthomonas Axonopodis ◽  
Sequence Characterized Amplified Region ◽  
Pcr Test

ABSTRACT Efficient control of Xanthomonas axonopodis pv. dieffenbachiae, the causal agent of anthurium bacterial blight, requires a sensitive and reliable diagnostic tool. A nested PCR test was developed from a sequence-characterized amplified region marker identified by randomly amplified polymorphic DNA PCR for the detection of X. axonopodis pv. dieffenbachiae. Serological and pathogenicity tests were performed concurrently with the nested PCR test with a large collection of X. axonopodis pv. dieffenbachiae strains that were isolated worldwide and are pathogenic to anthurium and/or other aroids. The internal primer pair directed amplification of the expected product (785 bp) for all 70 X. axonopodis pv. dieffenbachiae strains pathogenic to anthurium tested and for isolates originating from syngonium and not pathogenic to anthurium. This finding is consistent with previous studies which indicated that there is a high level of relatedness between strains from anthurium and strains from syngonium. Strains originating from the two host genera can be distinguished by restriction analysis of the amplification product. No amplification product was obtained with 98 strains of unrelated phytopathogenic bacteria or saprophytic bacteria from the anthurium phyllosphere, except for a weak signal obtained for one X. axonopodis pv. allii strain. Nevertheless, restriction enzyme analysis permitted the two pathovars to be distinguished. The detection threshold obtained with pure cultures or plant extracts (103 CFU ml−1) allowed detection of the pathogen from symptomless contaminated plants. This test could be a useful diagnostic tool for screening propagation stock plant material and for monitoring international movement of X. axonopodis pv. dieffenbachiae.

Sign in / Sign up

Export Citation Format

Share Document