| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2000: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1999: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Research Abstract |
HLA-DMB typing was performed from forensic samples (blood, bloodstain, hair, saliva stain, cerebral dura mater, urine, bone and tooth) using PCR-RFLP method. PCR products of the DMB allelic third exon was digested with three restriction endonucleases such as ApaL I, HinPl I, Bsr I.After digestion, samples were electrophoresed in 12% acrylamide gels. Most PCR products could be digested by those endonucleases and eight generic types were observed. Ninety-two percent of all the sample (N=129) was identified and eighty-three percent of the hair sample was identified. DMB^*0106 allele were not identified in this study. Although three pairs of heterozygous generic types could not distinguished each other, they could be typed by PCR-SSP method. DNA typing for HLA-DMB gene by this method could be available for the forensic applications such as personal identification, because of its reproducibility and simplicity.
DNA typing for DMB using PCR-SSP method was also studied. The problem of the false positive and false negative should be resolved. The screening of the allelic variations was also studied by PCR-SSCP method and two types were observed in the first exon. About another exon the screening will be performed and direct sequencing of the PCR products will be analyzed. The differentiation of the allelic variations was difficult by PCR-SSCP method and the primer design and the strategy should be reconsidered.
In this study, sample DNA was too little to analyze the DMB allelic sequence using direct sequencing method.
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