Transposon Tagging and Reverse Genetics
Transposons are mobile genetic elements that can amplify themselves in a genome. Transposable elements were first discovered in maize (McClintock 1948) and have been found to exist in all organisms. Transposons have multiple modes of movement or transposition, which is used to group the elements into two major classes, based on having an RNA or a DNA intermediate during transposition (reviewed in Hua-Van et al. 2005). The maize genome contains examples of most known transposon families including long terminal repeat (LTR) and non-LTR retrotransposons (class I), which use RNA intermediates, as well as class II DNA elements (Bruggmann et al. 2006). Maize class II elements tend to insert near or within genes (Bureau and Wessler 1992; Cowperthwaite et al. 2002; Fernandes et al. 2004; Kolkman et al. 2005; Kumar et al. 2005; McCarty et al. 2005; Settles et al. 2004). Transposition into genes can cause mutant phenotypes, and transposons are used as endogenous mutagens. This chapter focuses on the use of maize DNA transposons in molecular genetics and functional genomics studies.
With the exception of helitrons, DNA transposons share some common molecular and genetic properties. DNA elements have terminal inverted repeats as well as autonomous and non-autonomous transposons (Hua-Van et al. 2005). Autonomous elements encode the genes required for transposition. Non-autonomous transposons contain sequences recognized by transposase proteins and can move only in the presence of an autonomous element. Non-autonomous elements either have mutations in transposase genes or have replaced them with other sequences. DNA transposons also create target site duplications at the site of insertion. The length of the duplication is specific to each family of element. The major families of maize transposons that have been used as mutagens include Activator and Dissociation (Ac/Ds), Enhancer/Suppressor-mutator (En/Spm), and Robertson's Mutator (Mu). These families were identified because they cause unstable mutations (McClintock 1950, 1954; Peterson 1960; Robertson 1978). The instability of many transposon-induced alleles is due to excision events or epigenetic regulation of the transposons. The structures, mechanisms of transposition, and epigenetic regulation of Ac/Ds, En/Spm, and Mu are discussed in greater detail in multiple reviews (see Kunze and Weil 2002; Lisch 2002; Walbot and Rudenko 2002).
KeywordsLong Terminal Repeat Reverse Genetic Maize Genome Lateral Organ Boundary Domain Somatic Insertion
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