Difference between revisions of "Part:BBa K4729707"
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== The problem == | == The problem == | ||
− | == The virG Master-Switch construct == | + | === The virG Master-Switch construct === |
− | === Constitutive expression === | + | ==== Constitutive expression ==== |
=== Inducible expression === | === Inducible expression === |
Revision as of 16:53, 10 October 2023
Ptac + VirG TiBo542 N80D + repABCa
Contents
Introduction
Agrobacterium mediated transformation is one of the most prolific methods for plant engineering, by far the most common method for iGEM teams as well. However, working with plant Synbio is still far from a straightforward endeavor. This year, we set our sights on creating molecular tools that facilitate achieving successful transformation in non-model plant species, one of them is our Best Composite Part.
In nature, members of the genus Agrobacterium (Aphaproteobacteria) are soil-borne plant pathogens that integrate hormone producing genes in the host plant genome to cause growth deformities and tumors. This mechanism can be explored by substituting the oncogenes by genes of interest, thus integrating them in the plant chromosomes. The most well known strain, Agrobacterium tumefaciens, promotes crown-gall disease in most dicotyledonous plants, however, other strains such as Agrobacterium rhizogenes are also able to transform plant cells (Bahramnejad et al., 2019; Barton et al., 2018). Despite their wide use, Agrobacterium mediated plant transformation is still only well established for a handful of model organisms, which makes it hard for iGEM teams to engineer local species.
Part of the problem lies in the high specificity between plants and the necessary Agrobacterium strain for transformation. As we found out, getting hold of the correct Agrobacterium strain for a plant species (if one is even known) is extremely hard, reaching up to hundreds of Euros and months of shipping time. This is made even worse by the confusing and often conflicting nomenclature of existing strains (De Saeger et al., 2021).
Therefore, we created a composite part that aims to improve the efficiency and host range of Agrobacterium strains by leveraging the virG transcription factor as a Master-Switch plant transformation. With it, we hope to enable future teams to further extend the garden of plant projects in iGEM.
The molecular machinery of plant transformation
The problem
The virG Master-Switch construct
Constitutive expression
Inducible expression
Sodium Salicylate inhibits cell growth
Cross-talk assay
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 3840
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 4468
Illegal PstI site found at 3840 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1741
Illegal BamHI site found at 262
Illegal BamHI site found at 1917
Illegal BamHI site found at 2209
Illegal XhoI site found at 227
Illegal XhoI site found at 3480
Illegal XhoI site found at 3834
Illegal XhoI site found at 7176 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 3840
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 3840
Illegal NgoMIV site found at 111
Illegal NgoMIV site found at 2054
Illegal NgoMIV site found at 3894
Illegal NgoMIV site found at 4119
Illegal NgoMIV site found at 4224
Illegal NgoMIV site found at 5254
Illegal AgeI site found at 5733 - 1000COMPATIBLE WITH RFC[1000]