Transposon tagging for gene discovery:
Transposon mutagenesis in plants has becomean increasingly useful tool for gene discovery as well-marked, versatile transposons have been developed. Most of the transposon tagging systems that have been developed for plants other than maize are based on the maize
) transposable elementfamily.
Transposon tagging with maize transposable elements:
, isolated from maize,transposes in dicots as well as other monocots and has been used as the basis for transposon tagging systems in a variety of plants, including tobacco, tomato, flax and
dvantages of transposon mutagenesis:
ntroduction of the insertional mutagen can be separated from mutagenesis by controllingthe supply of transposase to the transposon.2.
Remobilization of the element provides a rapid means of establishing that a mutation iscaused by the insertion.3.
Because excision is imprecise, transposition of an element out of a gene often yieldsfurther mutations.4.
Transposons of the
family exhibit a marked preference for short-range transposition, potentially reducing the labor of identifying an insertion in a target gene by use of aclosely linked transposon.
Use of Transposable Elements - Transposon Tagging
- The transposable elements (TEs), in somecases, have been effectively utilized for isolation of genes, when the gene product is unknown. In this case atransposon works as a mutagen and therefore as a gene tag.Following steps which are involved in this procedure:(i) Clone a known gene with a scorable phenotypic effect.(ii) TE is transposed to this gene to get an unstable allele.(iii) This unstable allele is cloned and TE is isolated from this unstable allele (this is 10 select a TE whichcan produce unstable allele).(iv)This TE is transposed to a gene of interest with known phenotypic effect, to produce unstable allele.(v) The DNA is extracted from this mutant.(vi) TE sequence is used as a probe to isolate and clone the mutant gene (carrying inserted TE), so that wecan then isolate the gene of interest.In maize, TE like Ac/Ds, En/Spm and Mu 1 have been isolated using the genes Wx, C2 and Adh1. Similarly,TEs like Tam3 and Tam7 have been isolated from snapdragon (Antirrhinum majus). These TEs have beenused for gene tagging experiments leading to isolation of genes. In maize, several genes like Bz1, P, A1, Cl