HAGA Toshiaki TOKYO DENTAL COLLEGE,DEPARTMENT OF DENTISTRY,ASSIATANT, 歯学部, 助手 (80287185)
HANAOKA Yoichi TOKYO DENTAL COLLEGE,DEPARTMENT OF DENTISTRY,LECTURER, 歯学部, 講師 (30180912)
|Budget Amount *help
¥3,400,000 (Direct Cost : ¥3,400,000)
Fiscal Year 1998 : ¥1,500,000 (Direct Cost : ¥1,500,000)
Fiscal Year 1997 : ¥1,900,000 (Direct Cost : ¥1,900,000)
The purpose of this study is preparation of database on several DNA polymorphisms and their application to personal identification in the field of forensic dentistry and in the case of mass disaster. Differential PCR amplification of D12S67 locus resulted in increase in the number of discrete alleles from 11 to 35 and increase of heterozygosity from 80% to 93%. Comparison of nucleotide sequences of 188 alleles further increased the number of alleles to 52 and heterozygosity to 96%. Polymorphisms of D12S66, D21S11, VWA, DYS 19, and D4S43 loci were investigated using 170, 141, 155, 119 and 131 samples, respectively. The heterozygosity calculated for D12S66, D21S1, VWA and D4S43 were 57, 80, 79 and 81%, respectively.
The frequency of allele 15 was dominated in the Japanese population in DYS 19. New alleles with different nucleotide sequences from the known alleles were found in D21S11 and VWA loci, and the conformation of the nucleotide sequence of allele 14 was different from other alleles in the VWA locus. Allele 1 of the D4S43 locus was further classified into 4 alleles, resulting in the increase of heterozygosity of the locus. Above mentioned loci could successfully applied to the detection of polymorphisms using teeth as a source of DNA.
Comparison of the nucleotide sequences of D12S67, D21S11 and VWA loci in primates supported the known divergence models of primates and also showed some characteristics of typical mutation pattern of STR.Unknown regions of the X and Y amelogeniri genes were sequenced and new methods of sex determination using highly degraded DNA was established. Amplification of DNA was possible from teeth heat-treated at 200ﾟC.Detection of DNA from saliva-stained resin pieces was also possible from samples left at room temperature until 2 months. We also showed theoretically that three polymorphisms, D12S67, D21SI11 and VWA, could discriminate individuals in a mass disaster with victims of more than 1000.