中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2013年
1期
18-21
,共4页
贺淑婷%迟媛媛%蒲双双%刘建伟%温红玲%王志玉%宋艳艳%许洪芝%赵丽
賀淑婷%遲媛媛%蒲雙雙%劉建偉%溫紅玲%王誌玉%宋豔豔%許洪芝%趙麗
하숙정%지원원%포쌍쌍%류건위%온홍령%왕지옥%송염염%허홍지%조려
HIV-1%艾滋病痴呆综合征%基因多样性
HIV-1%艾滋病癡呆綜閤徵%基因多樣性
HIV-1%애자병치태종합정%기인다양성
HIV-1%AIDS dementia complex%Genes diversity
目的 研究艾滋病痴呆综合征(ADC)患者体内HIV-1 nef基因变异规律,为ADC发病机制的研究提供依据.方法 提取1例ADC病例尸检标本的外周(脾)和中枢神经系统(基底核、额叶灰质、脑膜、颞叶)共5个部位组织中的基因组DNA,PCR扩增HIV-1 nef基因,与pMD19-T克隆载体连接后,蓝白斑筛选挑取阳性菌落测序,序列经BLAST分析后,每个部位取5个序列,利用Bioedit、MEGA4等生物学软件进行基因距离、系统进化树以及同义/非同义替换值(ds/dn)等分析.结果 该ADC患者感染的是HIV-1 B亚型,与标准序列HXB2比较,该病例的HIV-1 nef基因存在变异,且外周组织与中枢神经系统各组织间以及中枢神经组织不同部位的基因变异不同,相同组织来源的nef序列基因距离较小,外周和中枢的nef基因距离差异较大,该病例H IV-1 nef基因在外周和中枢神经系统变异时均受到负向选择压力.结论 ADC患者体内HIV-1 nef基因存在多样性,且不同组织部位的基因变异不同,其导致的Nef蛋白某些氨基酸位点的改变对其生物学活性的影响有待进一步研究.
目的 研究艾滋病癡呆綜閤徵(ADC)患者體內HIV-1 nef基因變異規律,為ADC髮病機製的研究提供依據.方法 提取1例ADC病例尸檢標本的外週(脾)和中樞神經繫統(基底覈、額葉灰質、腦膜、顳葉)共5箇部位組織中的基因組DNA,PCR擴增HIV-1 nef基因,與pMD19-T剋隆載體連接後,藍白斑篩選挑取暘性菌落測序,序列經BLAST分析後,每箇部位取5箇序列,利用Bioedit、MEGA4等生物學軟件進行基因距離、繫統進化樹以及同義/非同義替換值(ds/dn)等分析.結果 該ADC患者感染的是HIV-1 B亞型,與標準序列HXB2比較,該病例的HIV-1 nef基因存在變異,且外週組織與中樞神經繫統各組織間以及中樞神經組織不同部位的基因變異不同,相同組織來源的nef序列基因距離較小,外週和中樞的nef基因距離差異較大,該病例H IV-1 nef基因在外週和中樞神經繫統變異時均受到負嚮選擇壓力.結論 ADC患者體內HIV-1 nef基因存在多樣性,且不同組織部位的基因變異不同,其導緻的Nef蛋白某些氨基痠位點的改變對其生物學活性的影響有待進一步研究.
목적 연구애자병치태종합정(ADC)환자체내HIV-1 nef기인변이규률,위ADC발병궤제적연구제공의거.방법 제취1례ADC병례시검표본적외주(비)화중추신경계통(기저핵、액협회질、뇌막、섭협)공5개부위조직중적기인조DNA,PCR확증HIV-1 nef기인,여pMD19-T극륭재체련접후,람백반사선도취양성균락측서,서렬경BLAST분석후,매개부위취5개서렬,이용Bioedit、MEGA4등생물학연건진행기인거리、계통진화수이급동의/비동의체환치(ds/dn)등분석.결과 해ADC환자감염적시HIV-1 B아형,여표준서렬HXB2비교,해병례적HIV-1 nef기인존재변이,차외주조직여중추신경계통각조직간이급중추신경조직불동부위적기인변이불동,상동조직래원적nef서렬기인거리교소,외주화중추적nef기인거리차이교대,해병례H IV-1 nef기인재외주화중추신경계통변이시균수도부향선택압력.결론 ADC환자체내HIV-1 nef기인존재다양성,차불동조직부위적기인변이불동,기도치적Nef단백모사안기산위점적개변대기생물학활성적영향유대진일보연구.
Objective To study the genetic diversity of HIV-1 nef genes from a patient with AIDS dementia complex (ADC),so as to research the amino acid variability and the pathogenesis of ADC.Methods The nef gene was amplified with PCR from genomic DNA which was extracted from spleen and different brain tissues(basal ganglia,frontal gray matter,meninges,temporal lobe)of a patient who died of ADC.PCR products were cloned into the pMD19-T vector,after transformation and selection by ampicillin and blue/white spotting.Five of positive clones were sequenced and confirmed with BLAST.HIV-1 nef sequences were processed with BioEdit and MEGA4 to do Neighbor-Joining tree,p-Distances,and values of ds/dn.Results The samples were all identified as HIV-1 B and genetic variation exists in HIV-1 nef gene isolated from different tissues compared with HXB2.In addition,part of the changes were different between periphery and brain.Conclusion Variations exist in the HIV-1 nef gene extracted from the ADC patient and the variations from peripheral and central nerve tissues were different,these variations may change the function of Nef,and it needs more research.
