摘要
目的 研究LLA新的等位基因HIA-A＊3113的分子机制。方法 样本DNA抽提采用PEL-FREEZ抽提试剂盒，利用PCR方法扩增先证者HIA-A基因的第1～8外显子，PCR产物直接经TOPOTA克隆转染到质粒载体中获得等位基因的单链，对所得克隆进行第2、3、4外显子双向测序分析。应用序列特异性引物PCR方法证实测序所发现的突变。结果 先证者样本克隆测序得到两个等位基因，其中一个等位基因为A＊2402，另一个经BLAST验证其为新的等位基因，新的等位基因序列已递交GenBank（DQ206619，DQ206620，DQ206621）。与最接近的A＊310102等位基因序列相比，新的等位基因在第3外显子上有1个核苷酸不同，第456位G→C，导致第128位氨基酸E→D。结论 该等位基因为新的HLA-A等位基因，被世界卫生组织HLA因子命名委员会正式命名为HIA-A＊3113。
Objective To investigate the genetic basis for HLA novel allele HIA-A ＊ 3113 in Chinese population. Methods DNA was extracted from peripheral blood by PEL-FREEZ DNA extraction kit. The amplifi- cation of HIA-A exons 1 - 8 of the proband was performed and the amplified product was cloned using TOPO TA cloning sequencing kit to split the two alleles apart. Strands of exons 2, 3 and 4 of the chosen colonies were se- quenced. The PCR-SSP was performed to confirm the mutations detected by sequencing. Results The sequencing results showed HL4-A alleles of the proband as A ＊ 2402 and the novel allele. The sequences of the novel allele have been submitted to GenBank（DQ206619, DQ206620, DQ206621）. BLAST analysis proved the novel allele with one nucleotide difference as HIA-A ＊ 310102 in exon 3 at nucleotide position 456G→C. This results in which shifting from E to D at codon 128. Conchtsion This allele is a novel allele and has been officially named A ＊ 3113 by the WHO Nomenclature Committee.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2007年第2期120-122,共3页
Chinese Journal of Microbiology and Immunology
基金
浙江省医药卫生科学研究基金项目（20032003）
