NASC Stock Detail Page
NASC ID: N2110789
Name: MAP-mCH-dOCRLdead x mCit-2xPH(PLC)
ABRC stock number: CS2110789
Description: Each 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 Col0 plants were transformed by dipping. Primary transformants (T1) were selected in vitro on glufosinate (basta) or Hygromycine. 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. This line is part of the set N2110797.
Donation Date: 2021-03-27
| Donated by: | Ecole Normale Supérieure de Lyon Yvon Jaillais |
Donor Number: iDePP control x PI(4,5)P2 marker UBQ10prom::GVG::6
Stock type: individual line
Material type: seed
Comment: Gene name: Drosophila melanogaster OCRL catalytic domain (168-509) with the D468G mutation, making it catalyticaly dead. mCHERRY. P24Y (NASC ID: N2105613): geneticaly encoded biosensor for PI(4,5)P2 fused with mCITRINE (YFP), localized at the plasma membrane, PLCdelta1 (Rattus norvegicus).
| Status | Price (£) |
|---|---|
| Available | £11.00 |
Germplasm Info
Seed type: Transgenic
Mutagen: T-DNA Insertion
Background: Col0
Segregation status: homozygous
Growth requirement: Induction upon dexamathasone (dex) treatment . No phenotype in the absence of dex and no specific growth condition needed in untreated plants.
Phenotype
This is a cross, which allows evaluating that removal of PI(4,5)P2 from the plasma membrane by iDePP is due to dOCRL catalytic activity. iDePP is a system for the inducible depletion of PI(4,5)P2 in Arabidopsis. It consists of the catalytic domain of Drosophila OCRL (dOCRL, a 5-phosphatase), fused at its N-terminus to a membrane anchoring peptide (MAP, myristoylatoion and Palmitoylation) and a mCHERRY for visualisation. iDePP is dex (dexamethasone) inducible and impacts the steady state of PI(4,5)P2 starting few hours after induction. The MAP-mCH-dOCRLdead is a control (catalyticaly inactive). This line is the result from a cross with the high affitnity PI(4,5)P2 sensor P24Y (NASC ID: N2105613), 2xPH(PLC).