CAJ | 학술논문

目的研究趋化因子SDF-1α转录子基因突变,分析突变发生频率,以及其对HIV感染的影响.方法研究对象包括居住在中国香港地区的278名HIV阴性健康中国人,49例HIV阳性白种人和13名高危未感染中国人.分别提取外周血单个核细胞基因组DNA和总RNA.建立PCR及逆转录PCR法,对SDF-1α启动子(promoter)、开放读码区(ORF)和3'端非编码区(3'UTR)基因进行序列分析.用人工引入点突变的错配引物双重PCR法对一新发现突变进行筛查,分析其在研究对象中的分布及意义.结果对包括13名高危中国人、49例白种人和38名随机选取的健康中国人在内的100份标本测序分析显示,在SDF-1α启动子及ORF区内未见任何突变;在SDF-1α3'UTR发现一"GA"插入式基因突变,命名为SDF-1-3'GA(+),在GenBank中的序列号为AY874118.该突变在3组研究对象中均有发生,在健康中国人中的频率为15.1%(84/556),但在13名有HIV接触而未感染的高危人群中发生频率为30.7%(8/26).结论与本身重要生理功能相对应,SDF-1α基因十分保守;在其3'UTR新发现突变SDF-1-3'GA(+)是否影响HIV感染,值得进一步研究.
목적 연구추화인자SDF-1α전록자기인돌변,분석돌변발생빈솔,이급기대HIV감염적영향.방법 연구대상포괄거주재중국향항지구적278명HIV음성건강중국인,49례HIV양성백충인화13명고위미감염중국인.분별제취외주혈단개핵세포기인조DNA화총RNA.건립PCR급역전록PCR법,대SDF-1α계동자(promoter)、개방독마구(ORF)화3'단비편마구(3'UTR)기인진행서렬분석.용인공인입점돌변적착배인물쌍중PCR법대일신발현돌변진행사사,분석기재연구대상중적분포급의의.결과 대포괄13명고위중국인、49례백충인화38명수궤선취적건강중국인재내적100빈표본측서분석현시,재SDF-1α계동자급ORF구내미견임하돌변;재SDF-1α3'UTR발현일"GA"삽입식기인돌변,명명위SDF-1-3'GA(+),재GenBank중적서렬호위AY874118.해돌변재3조연구대상중균유발생,재건강중국인중적빈솔위15.1%(84/556),단재13명유HIV접촉이미감염적고위인군중발생빈솔위30.7%(8/26).결론 여본신중요생리공능상대응,SDF-1α기인십분보수;재기3'UTR신발현돌변SDF-1-3'GA(+)시부영향HIV감염,치득진일보연구.
Objective To determine the polymorphism in CXC chemokine SDF-1α transcript and its effects on HIV infection.Methods Three groups of study subjects lived in Hang Kong were recruited:278 HIV-heahhy donors of Chinese origin.49 HIV+Caucasians and 13 Chinese with high risk behavior to HIV but kept uninfected.Genomic DNA and RNA were extracted from eripheral blood mononucleal cell. The PCR and RT-PCR reaction were set up accordingly.Sequence of the SDF-1α promoter,the open reading frame(ORF)and the 3'untranslate region(3'UTR)were analyzed.Two steps PCR reaction using two reverseprimers with mismatched nucleic acid were employed to screen the frequency of a novel mutation. Results equencing analysis from 100 subjects indicated that non mutation happened in tlle promoter and ORF of SDF-1α.A novel mutation Was detected from 3'UTR of SDF-1α.It is a "GA" insertion in "G" rich region near the stop code of SDF-1α.The mutation Was named as SDF-1-3’GA+and submitted to GenBank (AY874118).The mutation happened in three roups.with allele frequency of 15.1% in the healthy Chinese.of 30.7% in the high risk Chinese group.Conclusions our results confirm that SDF-1 genes arerelatively conserved.None noteworthy mutation is identified in the promoter and ORF regions of SDF-1α However.a novel mutation is identified from the 3'UTR of SDF-1α. It would be worthwhile to etermine effect of the novel mutation on HIV infection.