| ForwardThe gene of coden human leukocyte antigen (HLA) is located 6p21. Daus-set confirmed the first hunman leukocyte antige Mac in 1958,it show the beginning of HLA researching. More concentration on HLA system in forensic science lies on high polymorphism of HLA . There are many multiplex-alleles in each locus of HLA gene. What's more, they heridity co-dominant and associate randomly. Thus, higher genetic polymorphism can be formed. Since HLA polymorphisms is derived from single base difference and each base associate at random ,the number of HLA genotypes is over 10 . This provides an ideal genetic maker for personal identification and paternity testing. Single nucleiotide polymorphisms are single base pair positions in genome DNA at which different sequence alterative (allele) exit in nomal individuals in some population (s) , wherein the least frenqency allele has an abundance of 1 % or greater. But SNPs in HLA is much higher. On the grounds of evaluation, polymorphism distribution frequency of nu-cleioside in human genome is 0.08% -0.2%. However, polymorphism distribution frequency of nucleioside in HLA-I gene is highly 8. 6%. Thus, single nucleiotide polymorphisms in HLA become a new hotpoint at present in forensic science.Multiplex amplification detecting several SNPs loci at the same time is a great progress in PCR technique. By the method , multiple SNPs loci information can be gained at the same time,which may enhance the personal identification power especially when the material is of very low quantity. This is a ideal chatter for forensic usage. IMS-JST011107, IMS-JST011109 and IMS-JST011110 locus located in HLA-DRA are SNPs in JSNP database. HLA-DRA is one of HLA-II antigen in heavy chain gene, which owns three alleles and four exons. Thesethree SNPs loci are located between exon three and exon four. Japanese researcher firstly obtained allele frequency distribution data of HLA-DRA gene in Japanese group by genome scaning in 2000. In our country, there is still no genetic population data about them available. In this research, PCR-SSP combined with multiplex amplification was done to investigate the allele frequency distribution in Northern Chinese Han group. The data can serve as the base for the three SNPs loci's usage in forensic identification. The value of forensic application a-bout them was also evaluated.Materails and MethodsIn this study, IMS-JST011107JMS-JST011109 and IMS-JST011110 locus in HLA-DRA locus were investigated in 170 Northern Chinese Han group by u-sing PCR-SSP combined multiplex amplification method followed by polyacryl-amide gel electrophoresis( PAGE) and silver staining. The optimum condition of multiplex amplification in these loci was explored and applied in forensic identification.ResultsThe result of the multiplex amplification was perfect when annealing temperature was 58掳C and the ratio of primers concentration among IMS-JST011107> IMS-JST011109 and IMS-JST011110 in two amplification volumes were respectively 0.4/0. 6/1. 0( fjunol/L) and 0. 8/0. 8/0. 8(jjjnol/L).Three genotypes were detected in Northern Chinese Han group at IMS-JST011107 locus and two alleles,named T and C,whose frequency is respectively 0. 3029 and 0. 6973. These three genotypes are respectively T/T( 15) and whose frenqency is 0. 0882, C/T( 73 ) and whose frenqency is 0.4249, C/C (82 ) and whose frenquency is 0. 4823. Three genotypes were detected in Northern Chinese Han group at IMS-JST011109 locus and two alleles, named T and C, whose frequency is respectively 0. 3176 and 0. 6823. These three genotypes are respectively T/T (12) and whose frenqency is 0. 0706, C/T (84 ) and whosefrenqency is 0. 4941, C/C (74) and whose frenquency is 0. 4353. Three genotypes were detected in Northern Chinese Han group at IMS-JST011110 locus and two alleles, named A and C, whose frequency is respectively 0. 3088 and 0. 6912. These three genotypes are respectively A/A(11) and whose frenqency is 0. 0647, C/A (83 ) and whose frenqency is 0. 4882, C/C ( 76 ) and whose frenquency is 0.691...
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