MAP-BASED GENOME MINING METHOD FOR IDENTIFYING GENE REGULATORY LOCI

摘要

The invention pertains to a method for identifying one or more regions within a genome of an organism of interest that mediate the expression of one or more genes of interest The method comprises identifying a first and a second organism of interest, the first organism of interest is characterized by exhibiting a measurable response to an environmental stimulus, or otherwise exhibiting a phenotype associated with differential gene expression associated with a process of interest. The second organism of interest is characterized by lacking or not exhibit as strong a response to the stimulus as that observed within the first organism of interest, or it exhibits a different phenotype compared with that of the first organism of interest, wherein the different phenotype is associated with the process of interest or it exhibits a phenotype of interest that segregates when compared with the first organism of interest, or a combination thereof. The first and second organisms of interest are crossed to produce a population of segregated progeny and RNA is extracted from each of the segregated progeny. The level of gene expression for one or more genes of interest that are associated with the response to an environmental stimulus, or process of interest is quantified. A linkage map of the segregated progeny using one or more markers is prepared, and the relationship between said one or more markers on the linkage map and the gene expression of the one or more genes of interest is determined and one or more quantitative trail loci (QTL) identified. This method also pertains to identifying one or more QTLs associated with one or more genes of interest in segregated progeny that are subjected a desired environmental stimulus. Furthermore, this method may be used for the identification of one or more QTL corresponding to a transcription factor or any factor controlling the expression of one or more genes of interest, and the one or more genes located at the one or more QTL may be isolated and characterized.