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Biomedical subjects

A E Kwitek

Publications and source records attributed to A E Kwitek.

At least 19 recordsLinked to original sources
PubMed1993

The sensitivity of single-strand conformation polymorphism analysis for the detection of single base substitutions.

Single-strand conformation polymorphism (SSCP) analysis has proven to be a simple and effective technique for the detection of single base substitutions. We have used SSCP to analyze 29 mouse globin mutations, 27 p53 mutations, and 8 rhodopsin mutations contained within different size PCR products. Our results indicate that the type of mutation (transition versus transversion) did not play a major role in determining whether a mutation was detected by SSCP analysis. The position of the base substitution was more important than the precise base substitution in determining whether a mutation was detected. We report that SSCP sensitivity varies dramatically with the size of the DNA fragment being analyzed. The optimal size fragment for sensitive base substitution detection by SSCP is approximately 150 bp. Our results illustrate the need to keep the size of the PCR fragment small when performing SSCP to detect mutations. Larger fragments can be analyzed when screening for polymorphisms when the need to detect every sequence variation is not as critical.

Animals↗
PubMed1993

A denaturing gradient gel electrophoresis assay for sensitive detection of p53 mutations.

p53 is a tumor suppressor gene located on 17p, a region of the human genome frequently deleted in tumors. Mutation of the p53 gene is an important step leading to development of many forms of human cancer. To simplify the analysis of tumors for p53 point mutations, we describe a GC-clamped denaturing gradient gel assay for detecting single-base substitutions within highly conserved regions of the p53 gene. This assay allows for efficient screening of tumors for single-base substitutions within the p53 gene and can be used to facilitate sequence analysis of p53 point mutations.

Base Sequence↗
PubMed1993

Clinical features and linkage analysis of a family with autosomal dominant juvenile glaucoma.

BACKGROUND: Juvenile glaucoma is an uncommon form of open-angle glaucoma that is usually recognized during childhood or early adulthood and which often has a strong family history. METHODS: The authors clinically characterized a large multigeneration family with autosomal-dominant, juvenile-onset, open-angle glaucoma. Linkage analysis with short tandem repeat polymorphisms was used to evaluate the Rieger's syndrome locus as the site of the disease-causing mutation. RESULTS: Forty members of a family with a five-generation history of open-angle glaucoma were examined. Clinical data were available from an additional five individuals, three of whom were decreased. Older family members provided limited information about the visual history of five other deceased individuals in the first three generations. Fifty-nine people were at 50% risk of harboring the disease-causing mutation; and of these, 30 were affected with glaucoma by examination or by family history. All affected patients had an affected parent. The average age at diagnosis was 18 years (range, 8-30 years). Affected family members tended to be myopic but lacked other ocular or systemic abnormalities. The intraocular pressures (IOPs) of affected individuals were commonly more than 50 mmHg when they were first examined. Gonioscopy showed the angles to be open, with no abnormal pigmentation, iris processes, or embryonic tissue. Topical medications were initially effective in controlling IOP, but surgery was usually required for long-term pressure control. The Rieger's syndrome locus on chromosome 4q25 was excluded as the site of the disease-causing mutation. CONCLUSION: Juvenile open-angle glaucoma can occur as an autosomal dominant trait with high penetrance. Genetic linkage analysis of the family reported here has the potential to identify the chromosomal location of a glaucoma-causing gene. This gene is genetically distinct from the chromosome 4 locus that was recently associated with Rieger's syndrome.

Adolescent↗
PubMed1992

Linkage of Rieger syndrome to the region of the epidermal growth factor gene on chromosome 4.

Rieger syndrome is an autosomal dominant disorder of morphogenesis in which previous cytogenetic arrangements have suggested chromosome 4 as a candidate chromosome. Using a group of highly polymorphic short tandem repeat polymorphisms (STRP), including a new tetranucleotide repeat for epidermal growth factor (EGF), significant linkage of Rieger syndrome to 4q markers has been identified. Tight linkage to EGF supports its role as a candidate gene, although a recombinant in an unaffected individual has been identified. This study demonstrates the utility of using polymorphic STRP markers when only a limited number of small families are available for study.

Abnormalities, Multiple↗
PubMed1991

Linkage mapping of D21S171 to the distal long arm of human chromosome 21 using a polymorphic (AC)n dinucleotide repeat.

An (AC)n repeat within the anonymous DNA sequence D21S171 was shown to be highly polymorphic in members of the 40 Centre d'Etude du Polymorphisme Humaine (CEPH) families. Ten different alleles at this marker locus were detected by electrophoresis on polyacrylamide gels of DNA amplified by the polymerase chain reaction (PCR) using primers flanking the (AC)n repeat. The observed heterozygosity was 66%. PCR amplification of DNA from somatic cell hybrids mapped D21S171 to human chromosome 21, and linkage analysis localized this marker close to the loci CD18, PFKL, D21S113 and D21S112 in chromosomal band 21q22.3. In CEPH family 12 a de novo allele has been observed in a maternally derived chromosome.

Autoradiography↗
PubMed1991

Mapping of human chromosome 5 microsatellite DNA polymorphisms.

Thirteen moderately to highly informative microsatellite DNA polymorphisms based on (dC-dA)n.(dG-dT)n repeats were mapped to segments of human chromosome 5 using both linkage analysis and a panel of somatic cell hybrids which contained rearranged chromosomes. The markers were distributed throughout most of the length of the chromosome from the regions p15.3-p15.1 to q33.3-qter. Maps of the sites of meiotic recombination within the reference families proved particularly useful for the purpose of integrating new polymorphisms into the existing linkage map.

Base Sequence↗