ISSN:1059-7794    

DOI:10.1002/humu.1380020202

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

ISSN:1059-7794    

DOI:10.1002/humu.1380020203

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

ISSN:1059-7794    

DOI:10.1002/humu.1380020204

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

摘要:

AbstractTyrosinemia type I is an autosomal recessive inborn error of metabolism caused by deficiency of the enzyme fumaryl acetoacetate hydrolase (FAH, EC 3.7.1.2.). We have used reverse transcription and the polymerize chain reaction to amplify the peptide coding region of the FAH cDNA from four patients with tyrosinemia type I. Chemical mismatch cleavage analysis and DNA sequencing were utilized to determine mutant alleles in all cases. A French Canadian patient was homozygous for a splice error mutation in the 3′ portion of the gene. A second patient, from a consanguineous pedigree in Iran, had the identical splice alteration. The third patient has a missense mutation, changing valine to glycine in codon 166. And finally two nonsense mutations in codons 357 and 364 were found in the fourth patient. © 1993 Wiley‐Liss,

ISSN:1059-7794    

DOI:10.1002/humu.1380020205

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

摘要:

AbstractWe have previously reported linkage of a large Finnish family with the generalized (Köbner) type of epidermolysis bullosa simplex to chromosome 12q in the region containing the type II keratin gene cluster (Ryynänen et al., Am J Human Genet 49:978–984, 1991). In this study, we examined the possibility that keratin 5, the type II keratin expressed in the basal keratinocytes, harbors the mutation in this family. Nucleotide sequencing revealed a T‐to‐C transition within exon 7 of the keratin 5 gene in the affected individuals of the family, while the unaffected individuals showed no evidence of C. The presence of the T‐to‐C transition in the affected individuals was confirmed by restriction enzyme digestion analysis with Ncil endonuclease, as well as with PCR amplification of specific alleles (PASA) analysis. The PASA analysis also indicated that the mutated allele was not found among the 100 alleles tested within the general Finnish population indicating that the mutated allele is not a common polymhism. Furthermore, the mutated allele was not present in nine individuals representing three different EBS families of Finnish origin. The T‐to‐C transition at the nucleotide level resulted in substitution of a leucine by a proline at the amino acid level, and the substitution affected a leucine residue which was invariant among eight different human keratins in a highly conserved segment at the carboxy‐terminal region of the keratin 5 polypeptide. In analogy with previously elucidated mutations in keratins expressed in basal keratinocytes, it is highly probable that the leucine‐to‐proline substitution in the keratin 5 gene, which completely cosegregated with the clinical phenotype, is the underlying cause of the blistering tendency in this family with EBS of the generalized, Köbner type.

ISSN:1059-7794    

DOI:10.1002/humu.1380020206

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

摘要:

AbstractIn the course of analysing mutation in the factor IX gene from 200 haemophilia B patients in Sweden and the UK, we have identified one patient with a prepeptide missense mutation He has severe, antigen negative haemophilia, and complete analysis of his coding sequence reveals a single base transversion (A → T) causing substitution of isoleucine by asparagine at position −30. This change disrupts the hydrophobic core of the prepeptide, a feature which is required for secretion. Thus, haemophilia in this patient is caused by a failure to secrete factor IX from the hepatocytes. © 1993 Wiley‐Lis

ISSN:1059-7794    

DOI:10.1002/humu.1380020207

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

摘要:

AbstractFabry disease, an X‐linked recessive disorder of glycosphingolipid catabolism, results from lesions in the α‐galaciosidase A gene leading to deficient or absent activity of the lysosomal hydrolase. To facilitate the detection of rearrangements in this 14‐kb gene, a method was developed for the PCR amplification of all seven exons from genomic DNA in a single multiplex reaction. The entire coding region and all the intron/exon boundaries were amplified as four products. Application of this method permitted the detection of all five partial deletions previously identified by Southern analysis. This rapid method can be used to identify gene rearrangements in affected hemizygotes and determine heterozygosity for at risk females in families with Fabry disease. © 1993 Wiley‐

ISSN:1059-7794    

DOI:10.1002/humu.1380020208

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

摘要:

AbstractFrom 10 to 30% of lung carcinomas examined to date contain mutant K‐ras genes. We report here that the mutant‐allele‐specific amplification (MASA) method may be useful for detection of the K‐rasmutations in cells obtained from the sputum of patients with lung cancer. The PCR product from one of five patients revealed an alteration when mixed oligonucleotides representing variants of the second letter at codon 12 of this gene were used as 5′ primers, and further experiments showed a mutation of GGT (Gly) to GAT (Asp)at codon 12. The MASA system could also be applied to an examination of metastatic lung carcinomas, particularly from adenocarcinomas in colon and pancreas in which frequent K‐rasmutations are detected, and to mass‐screening for colorectal tumors using DNA isolated from feces as template. © 1993 W

ISSN:1059-7794    

DOI:10.1002/humu.1380020209

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

摘要:

AbstractA method is described for the detection of restriction fragment length polymorphisms (RFLPs) in single copy genes in mammalian cells using one 5′‐labelled oligonucleotide. This linear amplification (LA) method employs a single oligonucleotide as primer, which is extended byTaqDNA polymerase up to a restriction enzyme cleavage site. The products are arithmetically amplified by thermal cycling. The size of the products are determined by the sequence of the oligonucleotide and the position of the restriction enzyme cleavage site. Hence, an RFLP can be observed by measuring the size of the products. Polymorphisms which differ in size by a small number of base pairs, as are found in (CA)nrepeats, are especially suitable for analysis by the LA procedure since the products are run on DNA sequencing gels. A number of genes were examined by the procedure and all produced a satisfactory signal including a GC‐rich template. It is proposed that the LA method would be suitable for large‐scale genetic linkage analysis. The LA procedure has many advantages including the ability to multiplex signals under the same conditions, and lower cost since only one primer is needed. © 1993 Wiley

ISSN:1059-7794    

DOI:10.1002/humu.1380020210

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY

摘要:

AbstractThe tumor suppressor gene p53 is involved in predisposition to a variety of human cancers, including those from Li–Fraumeni cancer family syndrome patients. Studies of inheritance of p53 germline mutations require confirmation of the mutation in the tumors from family members. These studies as well as other retrospective studies of tumor specific mutations, are often hampered by a lack of available fresh or frozen tumor tissue samples for DNA extraction to confirm the suspected p53 mutation. Here we describe a simple technique for DNA isolation that permits mutational analysis of p53 from minimal amounts of paraffin‐embedded archival tissue samples. © 1993 Wiley‐Lis

ISSN:1059-7794    

DOI:10.1002/humu.1380020211

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1993

数据来源: WILEY