Home | About NASC | Address/Staff
Links | Ask a question | Help

NASC

The European Arabidopsis Stock Centre

SWELL Lines

Donated by

  • Yvon Jaillais Equipe Signalisation Cellulaire (SICE), UMR 5667 Laboratoire Reproduction et Développement des Plantes, Ecole Normale Supérieure de Lyon
  • Gregory Vert I2BC (Institute for Integrative Biology of the Cell), Centre National de la Recherche Scientifique (CNRS)

Click here to view all 216 of these lines.

Description

SWELL line seeds

The SWELL line is a collection of markers expressed in specific root tissues and that allows functional genomic in Arabidopsis thaliana, including tissue specific gene induction, nuclei purification by the INTACT method and multicolour imaging. The SWELL line collection is subdivided in four sets as follows:

  • SAND lines: Cell type specific markers that label the cytoplasm (yellow) with a 4xYFP tag (Set N2106367).
  • RED TIDE lines: Cell type specific markers that label the nucleus (red) with a H2B-2xCherry tag (Set N2106368).
  • BREAK lines: For cell type specific nucleus purification (green) with a NTF tag (INTACT). The BREAK lines are in a ACTIN2prom::BirA background (see Deal and Henikoff 2010 Dev Cell) (Set N2106369).
  • LINE-UP lines: for cell type specific inducible gene expression

SWELL line DNA

The SWELL line DNA collection is subdivided in three sets as follows:

  • Each SWELL promoter cloned into L4-L1R gateway compatible vectors (except the entry vectors for the following promoters: S4, S17, S18, S29, S32, IAA19, DR5, E47, UPB1 and Q12 that were provided by Philip Benfey (Duke University, NC, USA)). These entry vectors are kanamycin resistant in bacteria. (Set N2106366)
  • The destination vectors corresponding to each SAND (Set N2106363) , RED TIDE (Set N2106364) , BREAK (Set N2106365) and LINE-UP constructs. They are spectinomycin resistant in bacteria.
  • The various tag used in the Marquès-Bueno et al. paper cloned in their respective gateway compatible vectors (except the entry vectors for H2B that was provided by the Frederic Berger (GMI, Austria)).

Growth requirements in plants

  • All the SAND lines are Basta resistant
  • All the RED TIDE lines are Hygromycin resistant
  • All the BREAK lines are Kanamycin resistant (cell type specific NTF transgene) and basta resistant (BirA transgene)
  • All the LINE-UP lines are Basta resistant

Distribution

These seed and DNA stocks can all be ordered individually or as sets.

Individuals producing these lines

Maria del Mar Marquès-Bueno, Anne Cayrel, Matthieu Pierre Platre, Yvon jaillais and Grégory Vert.

Plant transformation

Each SWELL line construct was transformed into C58 GV3101 Agrobacterium strain and selected on YEB media (5g/L beef extract; 1g/L yeast extract; 5g/L peptone; 5g/L sucrose; 15g/L bactoagar; pH 7.2) supplemented with antibiotics (Spectynomicin, Gentamycin). After two days of growth at 28°C, bacteria were collected using a single-use cell scraper, re-suspended in about 200mL of transformation buffer (10mM MgCl2; 5% sucrose; 0.25% silweet) and plants (Col0 for SAND, RED TIDE and LINE-UP lines; BirA expressing Col0 plants for BREAK lines) were transformed by dipping.

SWELLINE lines selection and confirmation

Primary transformants (T1) were selected in vitro on the appropriate antibiotic/herbicide (glufosinate for SAND and LINE-UP lines, hygromycin for RED TIDE lines, and kanamycin for BREAK lines). For all constructs, around 20 independent T1 lines were isolated and six representative mono-insertion lines were selected in T2. Independent lines homozygous for the transgene were selected in T3. Some of the lines were produced later during the selection process and are distributed as segregating population.

Related Links

Further information for SWELL lines including localization images