【摘要】 目的了解RHD第9内含子碱基变异对其正常剪切是否存在影响。方法采用PCR方法扩增和测序分析中国汉族1名正常Rh阳性个体和2名DEL(D放散型)表型个体[携带RHD(1227A)等位基因]的RHD第9内含子,并做相互比对,同时通过NCBI Basic BLAST与参考序列(GenBank BN000065)加以比对。结果在3名中国汉族个体的第9内含子中,共观察到28处碱基变异(EMBL/GenBank/DDBJ EU372940～2),其中7处变异相同,可能为中国人第9内含子的特异性碱基变异;另有1处碱基变异仅在DEL样本中检出,在正常Rh阳性个体中未发现,可能为DEL特异性。结论未发现中国汉族人DEL第9内含子存在可能影响正常拼接的碱基变异,提示DEL mRNA缺失第9外显子相应的序列的分子机制即1227A>G碱基突变造成错误拼接所致。更多还原

【Abstract】 Subject To conduct sequencing analysis of RHD intron 9 in order to understand if any nucleotide variants interfered with intron splicing.Methods RHD intron 9 of an Rh D-positive and 2 DEL with RHD(1227A) allele individuals were sequenced via PCR amplifications. The sequences obtained were compared with each other and also compared with a reference EMBL/GenBank/DDBJ BN000065 sequence through NCBI Basic BLAST.Result Twenty-eight nucleotide variants were detected (EMBL/GenBank/DDBJ EU372940~2). Among them, seven variants were observed in all 3 samples, which may be the Chinese specific variants in RHD intron 9. One mutation was only found in the 2 DEL samples, but not in the normal D-positive, which may serve as DEL specific mutations. Conclusion Our study has not found any suspicious splicing-affecting variants in intron 9 of DEL allele, which may further prove that the molecular mechanism of whole exon 9 spliced out in DEL mRNA is no more than the 1227A>G mutation.更多还原