中华流行病学杂志
中華流行病學雜誌
중화류행병학잡지
CHINESE JOURNAL OF EPIDEMIOLOGY
2008年
11期
1119-1122
,共4页
郭俊涛%周艺彪%韦建国%赵根明
郭俊濤%週藝彪%韋建國%趙根明
곽준도%주예표%위건국%조근명
湖北钉螺%微卫星锚定聚合酶链反应%两翼序列
湖北釘螺%微衛星錨定聚閤酶鏈反應%兩翼序列
호북정라%미위성묘정취합매련반응%량익서렬
Oncomelania hupensis%Simple sequence repeat anchored polymerase chain reaction%Flanking sequence
目的 分析4个种群湖北钉螺中微卫星序列及其两侧序列的特点.方法 以(CA)8RY为引物对湖北钉螺基因组DNA进行微卫星锚定PCR(SSR-PCR)扩增,对全部159个扩增产物进行T克隆并测定其中82个片段的核苷酸序列.结果 SSR-PCR的扩增产物是湖北钉螺基因组DNA上的散在区域,不是微卫星序列,但扩增产物中含有微卫星序列,测序的82个片段中36个克隆片段含有微卫星序列.微卫星序列其侧翼序列有一定的保守性,同一个微卫星的侧翼序列多数情况下是相同的.(GA/CT)n、(TTAGGG/CCCTAA)n两类微卫星在4个钉螺种群中均有发现,(CAA)n仅在福建福清种群中发现,(TCTCTG)n仅在安徽贵池种群中发现,(GAA~TTC)n、(CAA/TTG)n、(CAT)n三种微卫星序列仅在四川普格种群中发现.结论 SSR-PCR扩增的不是微卫星,其结果的分析应当类似于随机扩增多态性DNA.因此SSR-PCR不能十分有效体现微卫星作为分子标记的优势,应当根据微卫星的侧翼序列设计针对微卫星的引物,对微卫星进行PCR扩增并进行分析.
目的 分析4箇種群湖北釘螺中微衛星序列及其兩側序列的特點.方法 以(CA)8RY為引物對湖北釘螺基因組DNA進行微衛星錨定PCR(SSR-PCR)擴增,對全部159箇擴增產物進行T剋隆併測定其中82箇片段的覈苷痠序列.結果 SSR-PCR的擴增產物是湖北釘螺基因組DNA上的散在區域,不是微衛星序列,但擴增產物中含有微衛星序列,測序的82箇片段中36箇剋隆片段含有微衛星序列.微衛星序列其側翼序列有一定的保守性,同一箇微衛星的側翼序列多數情況下是相同的.(GA/CT)n、(TTAGGG/CCCTAA)n兩類微衛星在4箇釘螺種群中均有髮現,(CAA)n僅在福建福清種群中髮現,(TCTCTG)n僅在安徽貴池種群中髮現,(GAA~TTC)n、(CAA/TTG)n、(CAT)n三種微衛星序列僅在四川普格種群中髮現.結論 SSR-PCR擴增的不是微衛星,其結果的分析應噹類似于隨機擴增多態性DNA.因此SSR-PCR不能十分有效體現微衛星作為分子標記的優勢,應噹根據微衛星的側翼序列設計針對微衛星的引物,對微衛星進行PCR擴增併進行分析.
목적 분석4개충군호북정라중미위성서렬급기량측서렬적특점.방법 이(CA)8RY위인물대호북정라기인조DNA진행미위성묘정PCR(SSR-PCR)확증,대전부159개확증산물진행T극륭병측정기중82개편단적핵감산서렬.결과 SSR-PCR적확증산물시호북정라기인조DNA상적산재구역,불시미위성서렬,단확증산물중함유미위성서렬,측서적82개편단중36개극륭편단함유미위성서렬.미위성서렬기측익서렬유일정적보수성,동일개미위성적측익서렬다수정황하시상동적.(GA/CT)n、(TTAGGG/CCCTAA)n량류미위성재4개정라충군중균유발현,(CAA)n부재복건복청충군중발현,(TCTCTG)n부재안휘귀지충군중발현,(GAA~TTC)n、(CAA/TTG)n、(CAT)n삼충미위성서렬부재사천보격충군중발현.결론 SSR-PCR확증적불시미위성,기결과적분석응당유사우수궤확증다태성DNA.인차SSR-PCR불능십분유효체현미위성작위분자표기적우세,응당근거미위성적측익서렬설계침대미위성적인물,대미위성진행PCR확증병진행분석.
Objective To analyze the sequence of microsatellite and the flanking sequence from four populations of Oncomelania hupensis. Methods We cloned 159 SSR-PCR amplification products of a commonly used primer, (CA)8RY, using O. hupensis genomie DNA as template, and sequenced 82 products Results The sequences obtained were novel O. hupensis genomic sequences but not repeat simple sequence. It was observed that 36 out of 82 clones contained microsatellites between priming sites.The flanking sequences of certain microsatellite were invariant. Both (GA/CT). and (TTAGGG/CCCAA)n were found in four populations of O. hupensis. However, (CAA)n were found only in O. hupensis from Fuqing,Fujian province and (TCTCTG), were found only in O. hupensis from Guichi,Anhui province and (GAA/TTC)n, (CAA/TTG)n, (CAT), were found only in O.hupensis from Puge,Sichuan province. Conclusion The results obtained by SSR-PCR should not be interpreted as the amplification of microsatellite loci, and analytical rules similar to those for Random Amplified Polymorphic DNA should be used. SSR-PCR could not make the most of the priority of microsatellite. It seems better to amplify the microsatellites with the primers designed on the basis of the flanking sequence.