摘要
目的建立HLA-DPB1基因第2～3外显子测序方法，并分析其多态性。方法根据HLA-DPB1基因序列，设计位点特异性引物扩增242名浙江汉族人群HLA-DPB1第2～3外显子序列，PCR扩增产物经双酶切后对第2和3外显子进行双向测序分析，采用Assign3．5＋SBT分析软件判读HLA-DPB1等位基因型。结果PCR扩增获得特异性的单一目的片段，其产物经直接测序后得到清晰的序列图。242名浙江汉族人群中共检测到18个HLADPB1等位基因，频率超过0．05的等位基因为DPB1＊05：01：01／135：0l（0．4112）、DPB1＊02：01：02（0．1901）、DPB1＊04：01：01（0．1136）以及DPB1＊02：02（0．0620），并确认1个新等位基因DPB1＊168：01。在第3外显子中发现9个多态性位点，其中517A〉T为本实验新检出的1个多态性位点。结论本实验建立的HLA～DPB1第2～3外显子测序方法是可行的，HLA-DPB1第3外显子存在一定多态性。
Objective To establish a polymerase chain reaction sequencing based typing （PCR-SBT） method for HLA-DPB1 exons 2 and 3, and to analyze their polymorphisms. Methods Based on the sequences of HLA-DPB1 loci, locus specific primers were designed and applied to amplify the target sequences encompassing the entire exons 2 and 3 of HLA DPBI. The amplification products were digested by enzymes and directly sequenced in both directions. The genotype was assigned by Assign 3. 5 ＋ SBT software. Results Specific target fragment was obtained with the PCR amplification, and good quality electropherogram was derived by direct sequencing. Among 242 individuals from Zhejiang Han population, 18 HLA-DPB1 alleles were detected. Aileles with a frequency of〉0.05 have included DPB1 ＊ 05：01：01/ 135：01 （0.4112）,DPB1＊ 02：01：02 （0.1901）, DPB1 ＊2 04：01：01 （0.1136） andDPB1 ＊ 02：02 （0.0620）. A novel HLA DPB1 ＊ 168：01 allele has also been identified. Nine polymorphism sites were founded in the exon 3 region, which included a new SNP site 517 A〉T. Conclusion The PCR SBT method for exons 2 and 3 of HLA DPBI is reliable, which allowed detection of poiymorphisms in exon 3 of the HLA-DPBI gene.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2015年第1期40-43,共4页
Chinese Journal of Medical Genetics
基金
浙江省医药卫生科学研究基金（2012KYA061、2013RCB003、2013ZDA007）
浙江省白然科学基金（LY14H100001）
