RHD第9内含子全长序列分析  

Complete sequencing of RHD intron 9

作　　者：邵超鹏[1] 李桢[1] 熊文[1] 周丹[1] 

机构地区：[1]深圳市血液中心深圳市输血医学研究所,广东深圳518035

出　　处：《中国输血杂志》2009年第5期354-357,共4页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金课题(编号:30670893);深圳市输血医学重点学科经费资助课题(编号:2008年度)

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。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 RI-ID（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 sampies, 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 susplcious 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.

关 键 词：RHD基因 内含子 序列分析 碱基变异 PCR 中国汉族 

分 类 号：R457.11[医药卫生—治疗学] Q503[医药卫生—临床医学]