The Plant Nuclear Marker Collection
Donated by
- Peter Shaw Department of Cell and Developmental Biology, John Innes Centre
- John Brown Division of Plant Sciences, College of Life Sciences, University of Dundee account (DEL0), James Hutton Institute
- Liam Dolan Department of Cell and Developmental Biology, John Innes Centre
Click here to view all 84 of these lines.
Description
The Plant Nuclear Marker collection contains a number of gateway constructs of Arabidopsis nuclear and nucleolar localised proteins. In most cases, the construct is available as an entry clone (pDONR207), GFP destination clone (GFP-N-BIN) and mRFP destination clone (pROK2-mRFP). These constructs are useful as markers of nuclear bodies and the nucleolus in co-localisation studies. These constructs have mostly been published in the papers referenced below. Images of many of the nucleolar markers are shown in the Plant Nucleolar Database (http://bioinf.scri.ac.uk/cgi-bin/atnopdb/home).
The donors would like to thank Ben Trewaskis for the gift of GFP-NBINOrdering
Clones can be ordered separately (see above link for list of all clones) or within the following sets:
| Name | NASC code | Sets Available | |
|---|---|---|---|
| pDONr207 Entry Clones | N794985 | Set of 28 clones | |
| GFP-N-bin Destination Clones | N794986 | Set of 24 clones | |
| pROK2-mRFP Destination Clones | N794987 | Set of 20 clones | 70 |
Seeds can be ordered individually and include the following lines:
| Name | NASC code |
|---|---|
| Coilin mRFP | N799456 |
| Fibrillarin GFP | N799457 |
| Eif4A-3 GFP | N799458 |
| cenH3 mRFP | N799459 |
| potato U2B" GFP | N799460 |
| pcb-1 | N799461 |
| ncb-1 | N799462 |
| ncb-2 | N799463 |
| ccb | N799464 |
Details of lines
cDNAs used for the generation of most of these constructs were available as trimmed U clones (cDNAs trimmed to the annotated start and stop codons in the Cre-Lox pUNI vector) from the SSP Orfeome collection (http://signal.salk.edu/SSP/). In other cases the cDNA was generated by PCR from an Arabidopsis thaliana cDNA library.
Coding regions with stop-codon deleted were amplified by high-fidelity PCR and then inserted by recombinase cloning into a standard Gateway entry vector (pDONR207; www.invitrogen.com).
Entry vector clones were transferred by recombinase cloning, into Gateway binary plant expression vectors GFP-N-BIN and pROK2-mRFP (which contains the coding sequence of monomeric RFP (Campbell et al., 2002) at the C-terminal end of the AttR cassette).
Agrobacterium mediated transformation was used to create GFP-NBIN and mRFP construct transformed lines. Mutant Arabidopsis thaliana lines ncb-1, ncb-2, pcb and ccb, as published in Collier et al, 2006 are also available.
References
Brown, J.W. et al. 2005. Arabidopsis nucleolar protein database (atnopdb). Nucleic Acids Research 33(Database Issue): D633-6. PMID. 15608277.
Campbell, R.E. et al. 2002. A monomeric red fluorescent protein. Proceedings of the National Academy of Sciences of the USA 99(12):7877–82. PMID. 12060735.
Collier, S. et al. 2006. A distant coilin homologue is required for the formation of Cajal bodies in Arabidopsis. Molecular Biology of the Cell 17(7): 2942-51.PMID. 16624863.
Kim, S.H. et al. 2009. Aberrant mRNA transcripts and the nonsense-mediated decay proteins upf2 and upf3 are enriched in the Arabidopsis nucleolus. Plant Cell 21 (7): 2045-2057.PMID. 19602621.
Koroleva, O.A. et al. 2009. Dynamic behavior of Arabidopsis eif4a-iii, putative core protein of exon junction complex: Fast relocation to nucleolus and splicing speckles under hypoxia. Plant Cell 21(5):1592-1606.PMID. 19435936
Pendle, A.F. et al. 2005. Proteomic analysis of the Arabidopsis nucleolus suggests novel nucleolar functions. Molecular Biology of the Cell 16(1):260-269.PMID. 15496452.