CAJ | 학술논문

目的通过对"亚洲型"DEL基因第7内含子全长序列测序分析,探讨其Mrna含有来自第7内含子170 bp片段的分子机制.方法根据美国国家生物信息中心(NCBI)GenBank BN000065的RHD基因序列,设计4对特异性寡核苷酸引物,分四段分别扩增RHD基因第7内含子.测序分析1名正常Rh阳性个体和2名RhDel表型个体(携带RHD1227A等位基因)的RHD第7内含子全长序列,然后通过NCBI Basic BLAST与参考序列(GenBank BN000065)进行比对,并作相互比对.结果发现3名个体第7内含子共观察到33处碱基变异(GenBank EU372940～2),其中8处变异相同;另有2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.结论这些变异不足以解释以往发现的DEL Mrna存在170 bp来自第7内含子序列而正常D阳性个体却不存在的现象,但测序结果提示该片段二侧的AG-GT可能参与这一分子事件的形成,因其正好与第7内含子二侧的GT-AG拼接位点协同将内含子一分为二,但具体机制可能十分复杂.
목적 통과대"아주형"DEL기인제7내함자전장서렬측서분석,탐토기Mrna함유래자제7내함자170 bp편단적분자궤제.방법 근거미국국가생물신식중심(NCBI)GenBank BN000065적RHD기인서렬,설계4대특이성과핵감산인물,분사단분별확증RHD기인제7내함자.측서분석1명정상Rh양성개체화2명RhDel표형개체(휴대RHD1227A등위기인)적RHD제7내함자전장서렬,연후통과NCBI Basic BLAST여삼고서렬(GenBank BN000065)진행비대,병작상호비대.결과 발현3명개체제7내함자공관찰도33처감기변이(GenBank EU372940～2),기중8처변이상동;령유2처감기변이부재DEL양본중검출,재정상Rh양성개체중미발현.결론 저사변이불족이해석이왕발현적DEL Mrna존재170 bp래자제7내함자서렬이정상D양성개체각불존재적현상,단측서결과 제시해편단이측적AG-GT가능삼여저일분자사건적형성,인기정호여제7내함자이측적GT-AG병접위점협동장내함자일분위이,단구체궤제가능십분복잡.
Objective To investigate the molecular mechanisms of DEL mRNA with one segment of 170 base pairs from intron 7,the complete nueleotide sequences of Asia DEL intron 7 was analyzed.Methods Four pairs of specific oIigonucleotide primer were designed to amplify and analyze the sequence of RHD intron 7 of one RHD-positive and two DEL with RHD(1227A)allele individuals separately in four sections,according to the RHD sequence of(NCBI)GenBank BN000065.The sequences obtained were aligned each other with a reference of GenBank BN000065 sequence by NCBI Basic BLAST.Results The results showed that 33 nucleotide variants were detected(EMBL/GenBank/DDBJ EU372940～2).Eight variants were observed in all samples whereas two variants were only found in DEL samples,but none in the normal D-positive among them.Conclusions The specific variants in DEL intron 7 cannot demonstrate that DEL mRNA has one segment of 170 base pairs sequence from intron 7,but it is not in normal D-positive individuals.The sequencing results,however,indicate that the AG-GT at both ends of the segment may be concerned with the molecular even.The AG-GT may cut intron 7 into two introns with its normal splicing site(GT-AG),but the molecular mechanisms may be complicate.