摘要
选择２７ 个识别序列为４ ｎｔ 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ＰＣＲＲＦＬＰ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 各等位基因核苷酸位点的变异系数， 发现ＤＱＢ１ 不同血清型间等位基因第２ 外显子核苷酸序列变化有一定规律． 研究为建立一个简捷准确的ＨＬＡ 分型方法提供了新的理论基础．
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
出处
《中山大学学报（自然科学版）》
CAS
CSCD
北大核心
1999年第5期61-66,共6页
Acta Scientiarum Naturalium Universitatis Sunyatseni
基金
国家杰出青年科学基金
广东省自然科学基金
