国际医学寄生虫病杂志
國際醫學寄生蟲病雜誌
국제의학기생충병잡지
INTERNATIONAL JOURNAL OF MEDICAL PARASITIC DISEASES
2014年
1期
20-24
,共5页
曹得萍%廖琳%陈达丽%陈建平
曹得萍%廖琳%陳達麗%陳建平
조득평%료림%진체려%진건평
中国利什曼原虫%前鞭毛体%系统发育学%核糖体小亚基RNA%细胞色素氧化酶亚单位Ⅱ
中國利什曼原蟲%前鞭毛體%繫統髮育學%覈糖體小亞基RNA%細胞色素氧化酶亞單位Ⅱ
중국리십만원충%전편모체%계통발육학%핵당체소아기RNA%세포색소양화매아단위Ⅱ
Chinese Leishmania isolates%Promastigote%Phylogeny%Small subunit ribosomal RNA%Kinetoplast cytochrome oxidase Ⅱ
目的 比较核糖体小亚基RNA(small subunit ribosomal RNA,SSUrRNA)和细胞色素氧化酶亚单位Ⅱ(cytochrome oxidaseⅡ,COXⅡ)这2种分子标志在中国利什曼原虫系统发育学分析中的不同. 方法 提取利什曼原虫各虫株核基因组和线粒体基因组,PCR方法扩增中国不同地区利什曼原虫SSUrRNA和COXⅡ基因,扩增产物送上海生工生物工程技术服务有限公司测序,序列碱基通过Clustal X软件比对,用DAMBE软件进行单倍型分析、Mega4软件计算遗传距离,Mrbayes3.1.2软件构建贝叶斯进化树. 结果 COXⅡ分析结果表明,流行于中国的利什曼原虫虫株未形成一个单系群;GS7和XJ771属于杜氏利什曼原虫复合体;10株利什曼原虫形成的6个单倍型(MHOM/CN/93/GS7,IPHUCN/77/XJ771,MHOM/CN/84/JS1和MGER/CN/60/GS-GER20除外)形成1个单系群;而SSUrRNA分析结果表明:11株利什曼原虫(IPHL/CN/77/XJ771,MH OM/CN/93/GS7,MRHO/CN/88/KXG-2,MHOM/CN/84/JS1,MGER/CN/60/GS-GER20除外)形成一个独立枝;江苏株JS1和热带利什曼原虫聚在一起,不与杜氏利什曼原虫聚在一起. 结论 SSUrRNA和COXⅡ得出的结果较一致,但COXⅡ基因在分析利仟曼原虫的系统发育关系时,可能是一个更可靠的遗传标志.
目的 比較覈糖體小亞基RNA(small subunit ribosomal RNA,SSUrRNA)和細胞色素氧化酶亞單位Ⅱ(cytochrome oxidaseⅡ,COXⅡ)這2種分子標誌在中國利什曼原蟲繫統髮育學分析中的不同. 方法 提取利什曼原蟲各蟲株覈基因組和線粒體基因組,PCR方法擴增中國不同地區利什曼原蟲SSUrRNA和COXⅡ基因,擴增產物送上海生工生物工程技術服務有限公司測序,序列堿基通過Clustal X軟件比對,用DAMBE軟件進行單倍型分析、Mega4軟件計算遺傳距離,Mrbayes3.1.2軟件構建貝葉斯進化樹. 結果 COXⅡ分析結果錶明,流行于中國的利什曼原蟲蟲株未形成一箇單繫群;GS7和XJ771屬于杜氏利什曼原蟲複閤體;10株利什曼原蟲形成的6箇單倍型(MHOM/CN/93/GS7,IPHUCN/77/XJ771,MHOM/CN/84/JS1和MGER/CN/60/GS-GER20除外)形成1箇單繫群;而SSUrRNA分析結果錶明:11株利什曼原蟲(IPHL/CN/77/XJ771,MH OM/CN/93/GS7,MRHO/CN/88/KXG-2,MHOM/CN/84/JS1,MGER/CN/60/GS-GER20除外)形成一箇獨立枝;江囌株JS1和熱帶利什曼原蟲聚在一起,不與杜氏利什曼原蟲聚在一起. 結論 SSUrRNA和COXⅡ得齣的結果較一緻,但COXⅡ基因在分析利仟曼原蟲的繫統髮育關繫時,可能是一箇更可靠的遺傳標誌.
목적 비교핵당체소아기RNA(small subunit ribosomal RNA,SSUrRNA)화세포색소양화매아단위Ⅱ(cytochrome oxidaseⅡ,COXⅡ)저2충분자표지재중국리십만원충계통발육학분석중적불동. 방법 제취리십만원충각충주핵기인조화선립체기인조,PCR방법확증중국불동지구리십만원충SSUrRNA화COXⅡ기인,확증산물송상해생공생물공정기술복무유한공사측서,서렬감기통과Clustal X연건비대,용DAMBE연건진행단배형분석、Mega4연건계산유전거리,Mrbayes3.1.2연건구건패협사진화수. 결과 COXⅡ분석결과표명,류행우중국적리십만원충충주미형성일개단계군;GS7화XJ771속우두씨리십만원충복합체;10주리십만원충형성적6개단배형(MHOM/CN/93/GS7,IPHUCN/77/XJ771,MHOM/CN/84/JS1화MGER/CN/60/GS-GER20제외)형성1개단계군;이SSUrRNA분석결과표명:11주리십만원충(IPHL/CN/77/XJ771,MH OM/CN/93/GS7,MRHO/CN/88/KXG-2,MHOM/CN/84/JS1,MGER/CN/60/GS-GER20제외)형성일개독립지;강소주JS1화열대리십만원충취재일기,불여두씨리십만원충취재일기. 결론 SSUrRNA화COXⅡ득출적결과교일치,단COXⅡ기인재분석리천만원충적계통발육관계시,가능시일개경가고적유전표지.
Objective To compare the difference between the analysis on the phylogenetic relationships of Leishmania isolates from different foci in China with partial fragment of small subunit ribosomal RNA (SSUrRNA) gene sequences or kinetoplast cytochrome oxidase Ⅱ (COX Ⅱ) gene sequences.Methods Genomic DNA and kinetoplast DNA were extracted from the promastigotes.The CO Ⅱ and SSUrRNA genes were amplified using respective primers.Products of PCR were sent to a commercial company for sequencing using the same PCR primer.Combined sequences matrix was aligned using Clustal X (1.8) with default settings and refined manually.Bayesian tree was constructed using Mrbayes-3.1.2.Results Leishmania isolates from different foci in China does not form a mono-phyletic group.The isolate MHOM/CN/93/GS7 and the isolate IPHL/CN/77/XJ771 belong to Leishmania donovani complex.Six haplotypes representing 10 Chinese Leishmania isolates(except isolates MHOM/CN/93/GS7,IPHL/CN/77/XJ771,MHOM/CN/84/JS1 and MGER/CN/60/GSGER20) form a monophyletic group (results from CO Ⅱ genes database).Eleven Chinese Leishmania isolates (except IPHL/CN/77/XJ771,MHOM/CN/93/GS7,MRHO/CN/88/KX G-2,MHOM/CN/84/JS1,MGER/CN/60/GSGER20) form a subclade,near to Leishmania tarentolae (results from SSUrRNA genes database); the isolate MHOM/CN/84/JS1 is sister of L.tropica representing L.tropica complex with high posterior probability (PP=1.00),instead of clustering with L,donovani complex or with other 10 Chinese isolates.Conclusions The analysis results with either SSUrRNA or COX Ⅱ seemed consistent.However kinetoplast cytochrome oxidase Ⅱ gene might be a more reliable genetic marker than SSUrRNA to analyze the phylogentic relationship of Chinese Leishmania isolates.
