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NASC

The European Arabidopsis Stock Centre

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)

Click here to view all 32 of these lines.

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.