NASC Stock Detail Page
NASC ID: N16549
Name: 35S::GA5 in ga5-1 background
ABRC stock number: CS16549
Description: The full length GA5 cDNA was cloned into pGEM-T Easy vector (Promega, Madison, WI) and subcloned as a XbaI-SacI fragment into pBI121 binary vector to replace the GUS gene, producing pBI121-GA5 (pBI-GA5). The construct was introduced into Agrobacterium tumefaciens strain C58C1 to transform GA4-sprayed ga5-1 Arabidopsis plants. Starting at 4-weeks and continuing weekly, ga5 plants that were to be transformed were sprayed with an aqueous solution of 10-4 M solution of GA4, containing 5 percent (v/v) ethanol. This step was necessary to overcome dwarfing and ensure fertility. Surface-sterilized T1 seeds of transformed plants were selected on MS medium supplemented with 50 microgram/ml kanamycin. Resistant seedlings were transferred to soil for continued growth and T2 seeds from several individual kanamycin-resistant plants were pooled. Successive generations (T3, T4) were harvested from plants selected on kanamycin.
Donation Date: 2009-09-03
| Donated by: | Michigan State University John Ohlrogge |
Donor Number: 35S::GA5 in ga5-1 background
Stock type: individual line
Material type: seed
| Status | Price (£) |
|---|---|
| Available | £11.00 |
Germplasm Info
Seed type: Transgenic
Background: Ler-0 (N62)
Growth requirement: Since the genotype was not determined, germinate seeds in MS plates containing 50 mg/L kanamycin; transfer seedlings to soil, keeping the pots covered for a few days to allow plants to adapt to the lower humidity environment
Phenotype
Dwarf phenotype of ga5-1 plants was rescued by the introduction of a wild-type copy of the gene in the 35S::GA5/ga5-1 transgenic plants.