Mehlmer Binary Plasmids
Donated by
- Norbert Mehlmer Molecular Cell Architecture and Trafficking Group, Botanik, LMU Biozentrum Martinsried, Department Biologie I, Fakultät für Biologie , Ludwig-Maximilians-Universität München (LMU)
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Description
Description
This collection contains a set of binary plasmids which targets the calcium sensor aequorin to different subcompartments of the plant cell. These constructs are aimed to generate transgenic Arabidopsis plants via agrobacterial transformation. The set includes constructs targeted to the cytosol, nucleus, inner surface of the plasma membrane, mitochondrial matrix, chloroplast stroma, as well as the inner and outer surface of the chloroplast envelope membranes.
Related publication: A toolset of aequorin expression vectors for in planta studies of subcellular calcium concentrations in Arabidopsis thaliana. Mehlmer N, Parvin N, Hurst CH, Knight MR, Teige M, Vothknecht UC. J Exp Bot. 2012 Feb;63(4):1751-61. doi: 10.1093/jxb/err406. Epub 2012 Jan 2.
Detailed description
These sets of binary plasmids is based on the pBin19 binary plasmid (1). The coding sequence of aequorin was fused to different targeting peptides or organellar proteins as a means to localise it to various cell compartments. All constructs furthermore consist of a YFP-aequorin fusion that allows easy verification of the correct sub-cellular localisation. There are in total four sets of plasmids. The expression cassette is driven by either the 35S or the Ubiquitin-10 promoter, while at the same time plant selection resistances against Basta and kanamycin are available for each construct.
The modular set-up of the plasmids further allows the easy replacement of targeting sequences to include other compartments.
The modified pBin19 plasmid
Contents of the set (N799855)
N799855 is the set of binary plasmids which targets the calcium sensor aequorin to different subcompartments of the plant cell. It contains the following :
| Nascode | Localisation | Promoter | Plant selection | Paper (ID) |
|---|---|---|---|---|
| N799856 | chloroplast | 35S | Basta | pBIN-CHYA(B) |
| N799857 | cytosol only | 35S | Basta | pBIN-CYA(B) |
| N799858 | IE chloroplast | 35S | Basta | pBIN-IEYA(B) |
| N799859 | mitochondria | 35S | Basta | pBIN-MYA(B) |
| N799860 | nucleus | 35S | Basta | pBIN-NYA(B) |
| N799861 | OE chloroplast | 35S | Basta | pBIN-OEYA(B) |
| N799862 | plasma membrane | 35S | Basta | pBIN-PMYA(B) |
| N799863 | chloroplast | Ubiquitin | Basta | pBINU-CHYA(B) |
| N799864 | cytosol only | Ubiquitin | Basta | pBINU-CYA(B) |
| N799865 | IE chloroplast | Ubiquitin | Basta | pBINU-IEYA(B) |
| N799866 | mitochondria | Ubiquitin | Basta | pBINU-MYA(B) |
| N799867 | nucleus | Ubiquitin | Basta | pBINU-NYA(B) |
| N799868 | OE chloroplast | Ubiquitin | Basta | pBINU-OEYA(B) |
| N799869 | plasma membrane | Ubiquitin | Basta | pBINU-PMYA(B) |
| N799870 | cytosol and nucleus | Ubiquitin | Basta | pBINU-YA(B) |
| N799871 | cytosol and nucleus | 35S | Basta | pBIN-YA(B) |
| N799872 | nucleus | Ubiquitin | Kanamycin | pBINU-NYA(K) |
| N799873 | chloroplast | 35S | Kanamycin | pBIN-CHYA(K) |
| N799874 | cytosol only | 35S | Kanamycin | pBIN-CYA(K) |
| N799875 | IE chloroplast | 35S | Kanamycin | pBIN-IEYA(K) |
| N799876 | mitochondria | 35S | Kanamycin | pBIN-MYA(K) |
| N799877 | nucleus | 35S | Kanamycin | pBIN-NYA(K) |
| N799878 | OE chloroplast | 35S | Kanamycin | pBIN-OEYA(K) |
| N799879 | plasma membrane | 35S | Kanamycin | pBIN-PMYA(K) |
| N799880 | chloroplast | Ubiquitin | Kanamycin | pBINU-CHYA(K) |
| N799881 | cytosol only | Ubiquitin | Kanamycin | pBINU-CYA(K) |
| N799882 | IE chloroplast | Ubiquitin | Kanamycin | pBINU-IEYA(K) |
| N799883 | mitochondria | Ubiquitin | Kanamycin | pBINU-MYA(K) |
| N799884 | OE chloroplast | Ubiquitin | Kanamycin | pBINU-OEYA(K) |
| N799885 | plasma membrane | Ubiquitin | Kanamycin | pBINU-PMYA(K) |
| N799886 | cytosol and nucleus | Ubiquitin | Kanamycin | pBINU-YA(K) |
| N799887 | cytosol and nucleus | 35S | Kanamycin | pBIN-YA(K) |
Cited in
Genetically encoded Ca2+ indicators: Properties and evaluation. Pérez Koldenkova V, Nagai T. Biochim Biophys Acta. 2013 Jan 22. pii: S0167-4889(13)00024-4. doi: 10.1016/j.bbamcr.2013.01.011.
The role of calcium in chloroplasts--an intriguing and unresolved puzzle. Rocha AG, Vothknecht UC. Protoplasma. 2012 Oct;249(4):957-66. Epub 2012 Jan 8.
References
- Mehlmer, N. et al. 2012. A toolset of aequorin expression vectors for in planta studies of subcellular calcium concentrations in Arabidopsis thaliana. Journal of experimental botany 63(4):1751-61. PubMed ID: 22213817.
- Bevan, M. 1984. Binary Agrobacterium Vectors for plant transformation. Nucleic Acids Research 12(22):8711-21.PMID. 6095209.