Ralstonia-GG-Kit
(Kit #
1000000142
)
Depositing Lab: Thomas Lahaye
The Ralstonia-GG-Kit contains a set of backbone vectors and Level I modules that can be used for assembly of Ralstonia solanacearum species complex gene knockout and complementation constructs based on the Golden Gate cloning techniques.
This kit will be sent as bacterial glycerol stocks in 96-well plate format.
Original Publication
A modular toolbox for Golden-Gate-based plasmid assembly streamlines generation of Ralstonia solanacearum species complex knockout strains and multi-cassette complementation constructs. Wu D, Schandry N, Lahaye T. Mol Plant Pathol. (2017) doi: 10.1111/mpp.12632. PubMed PMID: 29077245 .
Description
The Ralstonia-GG-Kit contains a set of backbone vectors and Level I modules that can be used for assembly of Ralstonia solanacearum species complex gene knockout and complementation constructs based on Golden Gate cloning techniques. LI modules includes R. solanacearum species complex specific promoters, N-terminal and C-terminal tags, reporter genes, terminators and resistance genes. This toolkit enables the simultaneous integration of up to five expression cassettes into a user-chosen site of the genome. In combination with the previously published plant golden gate toolkit (Binder, A, et al. (2014) ; Martin Parniske Lab plasmids), it also provides a straight forward way for scientists to express R. solanacearum effector proteins in plants.
Figure- Golden Gate (GG) based assembly of Rssc gene knockout constructs. For details, please see original image in publication from Wu et al., Molecular Plant Pathology (2017) doi: 10.1111/mpp.12632
How to Cite this Kit
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.
For your Materials and Methods section:
“The Ralstonia-GG-Kit was a gift from Thomas Lahaye (Addgene kit #1000000142).”
For your Reference section:
A modular toolbox for Golden-Gate-based plasmid assembly streamlines generation of Ralstonia solanacearum species complex knockout strains and multi-cassette complementation constructs. Wu D, Schandry N, Lahaye T. Mol Plant Pathol. (2017) doi: 10.1111/mpp.12632. PubMed PMID: 29077245 .
Ralstonia-GG-Kit - #1000000142
- Resistance Color Key
Each circle corresponds to a specific antibiotic resistance in the kit plate map wells.
- Inventory
Searchable and sortable table of all plasmids in kit. The Well column lists the plasmid well location in its plate. The Plasmid column links to a plasmid's individual web page.
- Kit Plate Map
96-well plate map for plasmid layout. Hovering over a well reveals the plasmid name, while clicking on a well opens the plasmid page.
Resistance Color Key
| Gentamicin | |
| Chloramphenicol and Gentamicin | |
| Ampicillin |
Inventory
| Well | Plasmid | Resistance |
|---|---|---|
| A / 1 | LI A-B phrpB | Gentamicin |
| A / 2 | LI A-B pEPS | Gentamicin |
| A / 3 | LI A-B pGala7 | Gentamicin |
| A / 4 | LI C-D GUS | Gentamicin |
| A / 5 | LI C-D Dest | Chloramphenicol and Gentamicin |
| A / 6 | LI D-E V5 | Gentamicin |
| A / 7 | LI E-F Tfd | Gentamicin |
| A / 8 | LI F-G Spec | Gentamicin |
| A / 9 | LI F-G Gent | Ampicillin |
| A / 10 | LI F-G Kan | Gentamicin |
| A / 11 | LI F-G Tet | Gentamicin |
| A / 12 | LI F-G Cml | Gentamicin |
| B / 1 | LI Rsc202375 Upstream | Gentamicin |
| B / 2 | LI Rsc203337 Downstream | Gentamicin |
| B / 3 | LI LacZ | Ampicillin |
| B / 4 | LII destination | Ampicillin |
| B / 5 | LIII 1-6 Amp RC | Ampicillin |