NASC Stock Detail Page
NASC ID: N2110795
Name: MAP-mCH-dOCRLdead x mCit-P4M
ABRC stock number: CS2110795
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 PI4P marker UBQ10prom::GVG::6UAS:
Stock type: individual line
Material type: seed
Comment: Gene name: Drosophila melanogaster OCRL catalytic domain (168-509), mCHERRY, P4M (NASC ID: N2107346) domain of SidM effector protein from Legionella pneumophila with with mCITRINE, PI4P biosensor.
| 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 control corss for the iDePP system showing that induction of inactive (dead) OCRL has no impact on PI4P. 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). It is the result from a cross with the PI4P sensor mCit-P4M (NASC ID: N2107346), and it allows to check that iDePP has no effect on PI4P accumulation at the plasma membrane.