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NASC

The European Arabidopsis Stock Centre

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 81 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-NBIN

Ordering

Clones can be ordered ordered separately (see above link for list of all clones) or within the following sets:

Name NASC code Sets Available Price £
pDONr207 Entry Clones N794985 Set of 28 clones 90
GFP-N-bin Destination Clones N794986 Set of 24 clones 80
pROK2-mRFP Destination Clones N794987 Set of 20 clones 70

Seeds can be ordered individually and include the following lines:

Name NASC code Price £
Coilin mRFP N799456 4
Fibrillarin GFP N799457 4
Eif4A-3 GFP N799458 4
cenH3 mRFP N799459 4
potato U2B" GFP N799460 4
pcb-1 N799461 4
ncb-1 N799462 4
ncb-2 N799463 4
ccb N799464 4

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.

pDONr207

GFP-N-bin