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Content archived on 2024-04-30

Activation tagging as a means of plant gene isolation

Objective



Though the majority of the worlds major crop plants can now be transformed, one of the most pressing problems we face is the relative lack of plant genes of agronomic worth currently available for transfer into novel host plants. This proposal centers on the use of "activation tagging" to isolate plant genes controlling the expression of genes encoding proteins with a function in either developmental, or biochemical pathways relevant to biotechnology. Activation tagging involves the use of a gene tag engineered to contain transcriptional enhancer sequences.
Following the insertion of such a tag into the plant genome, the expression of flanking sequences becomes deregulated, producing a dominant mutation. Notably, the dominant mutation produced allows a direct selection for a defined phenotype among the population of primary transformants. Hence, selection schemes can be devised for the expression of genes of agronomic potential. In addition to the transcriptional enhancers, the tag contains sequences derived from a bacterial plasmid, thus allowing rescue of tagged genes directly in bacteria with relative ease.
To date, activation tagging has been used successfully to create plant mutants whose cells are able to proliferate in the absence of auxin, or cytokinin, in the culture media, as well as mutants resistant to chemical, or metal, selection. This project involves the further development of this tagging strategy to isolate genes playing a role in a) shoot/root formation, b) control of transcription of enzymes of lipid biosynthesis, c) the plant`s response to pathogens and d) DNA recombination. To carry this out novel selection schemes have been devised including the use of transgenic plant lines engineered to contain markers which normally remain inactive in plant cells in the absence of transcriptional action of specific inducer genes. Tagging will be carried out followed by selection for expression of the marker gene constructs. Tagged genes able to activate expression of the marker genes will be rescued from the tagged lines, characterised and then reintroduced into novel hosts, linked to constitutive promoters so that they are ectopically expressed. The genes isolated in this programme are likely to produce agronomically relevant traits such as changes in patterns of plant growth, a faster and more efficient defensive response, the synthesis of lipids in novel tissues and a means to optimise gene targeting. Hence, the genes isolated will not only be of academic relevance, but also have biotechnological potential.

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Coordinator

MAX-PLANCK-GESELLSCHAFT ZUR FOERDERUNG DER WISSENSCHAFTEN E.V.
EU contribution
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Address
Carl-von-Linné-Weg 10
50829 KOELN
Germany

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