Arabidopsis Biological Resource Center

Description:

Transgenic line; 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. Homozygous for the mutation, heterozygous for the transgene.

Growth Requirement: Kanamycin resistant (50 mg/L); transfer seedlings to soil, keeping the pots covered for a few days to allow plants to adapt to the lower humidity environment

Marker: kanamycin

Background: Ler (Landsberg erecta)

ABRC Comment:

Format Shipped: 100 seeds per vial

Base / Commercial Price: $15 / $120