HLA DQ位点多样性模拟分析及其PCR—RFLP分型

选择２７个识别序列为４ｎｔ的限制性内切酶，通过计算机对最新获得的１９个ＤＱＡ１和３５个ＤＱＢ１等位基因第２外显子中的一段序列进行模拟酶切分析，根据酶切格局数目及各格局所代表的等位基因数的均衡性，确定酶的优先次序，在此基础上进行ＰＣＲ－ＲＦＬＰ模拟分析，确定最佳的酶组合。结果表明，分别采用４个和６个酶的组合，可有效鉴定ＤＱＡ１和ＤＱＢ１等位基因。同时通过比较分析ＤＱＢ１各等位基因核苷酶位点的变异系数

Computer Analysis of Human HLA DQ Locus Polymorphism and Genotyping by PCR RFLP

The exon 2 sequences of 19 DQA1 and 35 DQB1 alleles bounded respectively on two pairs of primers were cleaved successively with a set of 27 restriction endonucleotidases with 4nt recognition sequence by using a new computer program in order to determine the enzyme cleaving pattern and select proper enzymes for genotyping The genotyping of DQA1 and DQB1 alleles by polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) was also performed with computer program Results demonstrate a set of 4 and 6 restriction endonucletase have efficiently genotyped DQA1 and DQB1 alleles by PCR RFLP, respectively Some alleles can be identified with other restriction endonucletase besides above 27 enzymes The analysis of polymorphism of DQB1 alleles suggest that variability of nucleotide sequences of exon II among DQB1 alleles of different serotypes presents some rules The study provide the theoretical base for DQB1 and DQA1 genotyping by PCR RFLP