ABSTRACT
Objective: To establish real-time qPCR method to analyze each HLA-C allele expression level of individual.Methods:Database including exon 2&3 sequences of HLA class Ⅰalleles was built ,HLA-C allelic specific primers were designed and the real-time quantitative PCR method for analyzing HLA-C allele expression level was built .The allelic specificity of these primers were confirmed in database and 835 normal peripheral blood samples of Han population .The mRNA level of each HLA-C allele from 20 pairs of liver tumor tissues and non-tumor tissues was analyzed by the qPCR method we built .Results:20 pairs of allelic specific primers were designed to distinguish the two HLA-C alleles of each individual with frequency over 0.96%in 835 cases of Han population.Among 55%of the liver cancer tissues ,the expression levels of the two HLA-C alleles from the same tumor tissue changes differently compared to that of the relevant non-tumor tissue.Conclusion:This study provides method for HLA-C allele expression level analysis of Han population and each HLA-C allele expression level is inconsistent of liver cancer tissue .
ABSTRACT
Objective:To establish a bidirectional amplification of specific alleles polymerase chain reaction(Bi-PASA PCR)technique system and to apply this technique to study of single nucleotide polymorphism(SNP)of genes.Methods: The information of nucleotide sequences and their corresponding SNPs of gene were obtained from genBank of National Central of Biotechnology Information.The primers were designed by Primer 5.0 software and their specificity was tested by NCBI Blast 2.0 software.Results: They successfully designed and established the technique system of Bidirectional amplification of specific alleles Bi-PASA.The method were applied to study SNPs of PTEN(phosphatase and tensin homology deleted on chromosome ten,PTEN).The superiority and stability of Bi-PASA PCR technique on determining SNPs of genes have been tested.Conclusions:The Bi-PASA PCR technique is a new method for determination of SNP with more convenience,more specificity,less cost and easy availably and has much more advantage especially in the study of population gene polymorphism.