Pilot study of arbitrarily primed PCR-single stranded DNA conformation polymorphism analysis for screening genetic polymorphisms related to specific phenotypes.
BACKGROUND
To investigate relationships between phenotypes and genotypes is not simple. We propose a phenotype-to-genotype screening strategy and pooled DNA system. As a pilot study of this strategy, we used arbitrarily primed polymerase chain reaction (AP-PCR) in combination with single-stranded DNA conformation polymorphism (SSCP) to screen for genetic polymorphisms associated with longevity.


METHODS
Study subjects were separated into 3 age groups, individuals aged >100 years, 90-99 years and 60-69 years. Genomic DNAs were prepared from each individual, pooled to represent the 5 study groups, and then the pooled genomic DNAs were subjected to AP-PCR-SSCP analysis.


RESULTS
We found 1 SNP more frequently in senior citizens with longevity. The genotype frequency of the 82133G>A polymorphism of human chromosome 3 clone RP11-61K12 (AC011199) differed significantly (P=0.0189, Fisher's exact test) between older subjects (>90 years) and younger subjects (<70 years). It is noteworthy that the strategy we describe herein was useful for identifying an SNP that showed statistically significant differences in its distribution across the subject groups.


CONCLUSIONS
The pooled DNA strategy and quantitative genotype discrimination can also be applied to screening for the relationship between phenotype and genotype more effectively.
Maekawa
Masato
M
Department of Laboratory Medicine, Hamamatsu University School of Medicine, 1-20-1 Handayama, Hamamatsu 431-3192, Japan. mmaek@hama-med.ac.jp
Taniguchi
Terumi
T
Uramoto
Takeshi
T
Higashi
Hitomi
H
Horii
Toshinobu
T
Takeshita
Akihiro
A
Sugimura
Haruhiko
H
Kanamori
Masao
M
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008136:Longevity / Q000235:genetics*; D008875:Middle Aged; D009690:Nucleic Acid Conformation; D010641:Phenotype; D010865:Pilot Projects; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic
D004277:DNA, Single-Stranded