Chapter title |
T-DNA Insertional Mutagenesis and Activation Tagging in Medicago truncatula
|
---|---|
Chapter number | 6 |
Book title |
Functional Genomics in Medicago truncatula
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8633-0_6 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8632-3, 978-1-4939-8633-0
|
Authors |
Francesco Panara, Ornella Calderini, Andrea Porceddu, Panara, Francesco, Calderini, Ornella, Porceddu, Andrea |
Abstract |
The development of plant genetic transformation techniques has greatly enhanced our capacity to investigate and understand gene function. Since T-DNA constructs insert randomly in genomes, in principle, it is possible to construct a population of individuals harboring one or more T-DNA inserted in any region of the genome. Such populations can be screened following two approaches: (1) given a mutant phenotype, one could find the gene subtending the phenotypic alteration (forward approach), or (2) given a gene of interest, one could identify the phenotypic effect of its expression perturbation (reverse approach).Activation tagging is an application of T-DNA mutagenesis aimed at obtaining gain-of-function mutations. This can be achieved by introducing enhancer sequences randomly in the target genome via a T-DNA shuttle and then analyzing the genomic regions flanking the insertion sites in individuals showing phenotypic alterations. In this chapter, we describe the detailed procedure to obtain and screen an activation-tagged population in Medicago truncatula. |
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