Plant transformation vector: Difference between revisions

Revision as of 17:53, 14 January 2015

Revision as of 20:19, 27 September 2015

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==Steps in plant transformation==

Propagate binary vector in ''E. coli''

Isolate binary vector from ''E.coli'' and engineer (introduce a foreign gene)

Re-introduce engineered binary vector into ''E. coli'' to amplify

Isolate engineered binary vector and introduce into ''Agrobacteria'' containing a modified (relatively small) Ti plasmid

Infect plant tissue with engineered ''Agrobacteria'' (T-DNA containing the foreign gene gets inserted into a plant cell genome)

In each cell T-DNA gets integrated at a different site in the genome

Note: There are many variations to these steps. A custom DNA plasmid sequence can be created and replicated in more than one way.

=== Consequences of the insertion ===

Foreign DNA inserted

Insertional mutagenesis (but not lethal for the plant cell – as the organism is diploid)