CAJ | 학술논문

目的研究人巨细胞病毒( human cytomegalovirns,HCMV) UL131 A-128序列在临床低传代分离株中的转录方式.方法用3'RACE技术扩增HCMV不同临床株,不同时期的UL131A,UL128,UL130的mRNAs并测序分析；利用PCR技术在HCMV临床株cDNA文库中筛选分别含有UL13IA,UL130,UL128序列的阳性克隆,测序并分析结果.结果成功在临床低传代分离株中得到UL13IA,UL128,UL130基因的转录结构,UL131A,UL130的转录方式和文献报道一致,UL131A含有两个外显子,UL130为此区域内唯一不含内含子的基因.而UL128基因的转录本呈现了两种转录方式,一种结构包含三个外显子,长度为519 bp；而另一种结构包含三个外显子和第一内含子结构,长度为642 bp,在不同病毒株的不同时期均显示了包含第一内含子结构转录本的含量明显高于仅有外显子结构的UL128转录本的含量.UL131A-128三个基因在病毒的即刻早期,早期,晚期均存在.3'RACE得到结果与HCMV cDNA文库筛选得到的结果相一致.结论 UL131A,UL130和UL128基因的mRNA结构的起始位点不同,但他们在3’末端拥有共同的终止位点.HCMV临床株UL 131A-128基因转录方式的复杂结构可能在HCMV感染的不同时期具有重要作用.
목적 연구인거세포병독( human cytomegalovirns,HCMV) UL131 A-128서렬재림상저전대분리주중적전록방식.방법 용3'RACE기술확증HCMV불동림상주,불동시기적UL131A,UL128,UL130적mRNAs병측서분석；이용PCR기술재HCMV림상주cDNA문고중사선분별함유UL13IA,UL130,UL128서렬적양성극륭,측서병분석결과.결과 성공재림상저전대분리주중득도UL13IA,UL128,UL130기인적전록결구,UL131A,UL130적전록방식화문헌보도일치,UL131A함유량개외현자,UL130위차구역내유일불함내함자적기인.이UL128기인적전록본정현료량충전록방식,일충결구포함삼개외현자,장도위519 bp；이령일충결구포함삼개외현자화제일내함자결구,장도위642 bp,재불동병독주적불동시기균현시료포함제일내함자결구전록본적함량명현고우부유외현자결구적UL128전록본적함량.UL131A-128삼개기인재병독적즉각조기,조기,만기균존재.3'RACE득도결과여HCMV cDNA문고사선득도적결과상일치.결론 UL131A,UL130화UL128기인적mRNA결구적기시위점불동,단타문재3’말단옹유공동적종지위점.HCMV림상주UL 131A-128기인전록방식적복잡결구가능재HCMV감염적불동시기구유중요작용.
Objective To study and research the transcription pattern of UL131A-128 mRNA in human cytomegalovirus (HCMV) clinical low passage strains.Methods The UL131 A-128 mRNAs of from different clinical strains and kinetic periods were amplified using 3 ' RACE and analyzed by sequencing.Meanwhile,clones containing UL131A-128 transcripts in a HCMV cDNA library of clinical strain were selected and sequenced.Results It is successful to obtain the transcription pattern of UL131 A,UL130 and UL128 gene in HCMV clinical low passage strains,the UL131A gene consisted of two exons and the coding region of UL130 gene was not interrupted by any intron in the region as reported before.However,the transcript of UL128 gene showed two patterns,one pattern consisted of the three exons,the length is 519bp,and the other one contained the three exons and the sequence of the first intron further,the length is 642bp.The quantities of UL128 transcript containing the sequence of the first intron were higher than that of transcript only containing the three exons in the studied clinical strains at all kinetic classes.It was demonstrated that the UL131A-128 mRNA were expressed with immediately early,early and late kinetics.The result of 3'RACE and HCMV cDNA library of clinical strain is conformity.Conclusions Our results demonstrated that the UL1 31 A,UL130 and UL128 genes were transcribed with 3'-coterminal,although the initiation points of their mRNA may be different.The variation of the transcripts found in our study indicated complex nature of transcription of UL131A-128 genes in HCMV clinical strains.