RESUMO
OBJECTIVE@#To set up the method for analyzing HLA-B gene polymorphism with PCR-RFLP, and to gain population data among northern Chinese Hans of HLA-B's restricted fragments after NlaIII digestion, and to achieve application in forensic medicine practice.@*METHODS@#Sample DNA was extracted by the phenol/chloroform extraction method, 943 bp-long fragments containing HLA-B exon 2 and 3 were got by PCR. The endonuclease NlaIII was applied to cut the PCR products into polymorphic fragments shorter than 943bp, then PAGE and silver staining were used to detect the digestion results, finally the digestion sites were assured by DNA sequencing.@*RESULTS@#Along 943bp-long PCR products, 14 length-different fragments, 20 kinds of fragment combinations were got and 6 cutting site were observed after NlaIII digestion.@*CONCLUSION@#HLA-B gene was highly polymorphic among Chinese northern Hans. Even with only one endonuclease, 14 restricted fragments were got and the PIC was great. Such a HLA-B PCR-RFLP analysis will have values in forensic medicine applications.
Assuntos
Humanos , Povo Asiático/genética , Sequência de Bases , China/etnologia , DNA/isolamento & purificação , Éxons , Medicina Legal/métodos , Frequência do Gene , Antígenos HLA-B/genética , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de RestriçãoRESUMO
OBJECTIVE@#To establish a method which processes RHCE genotyping with PCR.@*METHODS@#Using PCR-SSP to detect RHCE genotype in 200 cases of Han population in north of China and Li population in south of China and detecting 5 samples of parentage testing at the same time.@*RESULTS@#The results of RHCE genotyping in individuals of two populations are completely accorded with the results of serology typing. And the distribution of RH genotypic frequency in Han population in north China is: RHCCEE 1, RHCCEe 3, RHCCee 88, RHCcEE 4, RHCcEe 20, RHCcee 54, RHccEE 1, RHccEe 22, RHccee; The distribution of RH genotypic frequency in Li populatin in south China is RHCCEE 2, RHCCEe 2, RHCCee 106, RHCcEE 7, RHCcEe 62, RHCcee 10, RHccEE 3, RHccEe 8. The results of RHCE genotype detecting of parentage testing samples are accorded with the results of associated identification of 13 STR loci.@*CONCLUSION@#PCR-SSP technology can exactly detect RHCE genotype in individuals of Han population in north China and Li population in south China.