Cloning quantitative trait loci by insertional mutagenesis.

摘要: This study explores the theoretical potential of “insertional mutagenesis” (i.e., mutagenesis as a result integration novel DNA sequences into germ line), means cloning quantitative trait loci (QTL). The approach presented is based on direct search for mutagenic effects nature, and makes no assumptions to nature affecting value. Since there are very large number insertion sites in genome but only limited target that can affect any particular trait, numbers inserts will have be generated screened. allelic variants at single QTL phenotype value expected small relative sampling variation. Thus two three stages replicate testing required each insert order bring overall Type I error down negligible proportions yet maintain good statistical power with true traits under consideration. effort involved depend spectrum produced by insertional mutagenesis. presently unknown, using reasonable estimates, about 10,000 would tested, (n ≧ 30) (α=0.01) first stage, provide high likelihood detecting least one effect given interest. Total offspring per detected decrease strongly increased scored initial transformed parent. In fact, it appear successful implementation experiments this sort require introduction multiple independent original parent individuals, repeated transformation, or use transposable elements inserts. When biologically feasible, selfing method choice replication, all cases must carried out inbred lines reduce variation due genetic segregation, avoid confounding mutational linkage nearby segregating QTL. special qualifications Arabidopsis thaliana studies emphasized, problems raised somaclonal discussed.