Ayyadevara S, Ayyadevara R, Hou S, Thaden J J, Shmookler Reis R J
Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Genetics. 2001 Feb;157(2):655-66. doi: 10.1093/genetics/157.2.655.
Recombinant-inbred populations, generated from a cross between Caenorhabditis elegans strains Bergerac-BO and RC301, were used to identify quantitative trait loci (QTL) affecting nematode longevity. Genotypes of young controls and longevity-selected worms (the last-surviving 1% from a synchronously aged population) were assessed at dimorphic transposon-specific markers by multiplex polymerase chain reaction. The power of genetic mapping was enhanced, in a novel experimental design, through map expansion by accrual of recombinations over several generations, internally controlled longevity selection from a genetically heterogeneous, homozygous population, and selective genotyping of extremely long-lived worms. Analysis of individual markers indicated seven life-span QTL, situated near markers on chromosomes I (tcbn2), III (stP127), IV (stP13), V (stP6, stP23, and stP128), and X (stP41). These loci were corroborated, and mapped with increased precision, by nonparametric interval mapping-which supported all loci implicated by single-marker analysis. In addition, a life-span QTL on chromosome II (stP100-stP196), was significant only by interval mapping. Congenic lines were constructed for the longevity QTL on chromosomes III and X, by backcrossing the Bergerac-BO QTL allele into an RC301 background with selection for flanking markers. Survival data for these lines demonstrated consistent and significant effects of each QTL on life span.
利用秀丽隐杆线虫品系Bergerac - BO和RC301杂交产生的重组近交群体,来鉴定影响线虫寿命的数量性状基因座(QTL)。通过多重聚合酶链反应,在双态转座子特异性标记处评估年轻对照和长寿选择线虫(来自同步老化群体中最后存活的1%)的基因型。在一个新颖的实验设计中,通过几代积累重组来扩展图谱、从遗传异质的纯合群体中进行内部对照的长寿选择以及对极长寿线虫进行选择性基因分型,增强了基因定位的能力。对单个标记的分析表明有7个寿命QTL,位于染色体I(tcbn2)、III(stP127)、IV(stP13)、V(stP6、stP23和stP128)和X(stP41)上的标记附近。通过非参数区间定位对这些基因座进行了确证,并更精确地进行了定位,该方法支持单标记分析所涉及的所有基因座。此外,染色体II上的一个寿命QTL(stP100 - stP196)仅通过区间定位才具有显著性。通过将Bergerac - BO QTL等位基因回交到RC301背景中并选择侧翼标记,构建了染色体III和X上长寿QTL的近等基因系。这些品系的存活数据表明每个QTL对寿命都有一致且显著的影响。