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Abstract This study was designed to optimize the factors about onion (Allium cepa) epidermal cells infected by Agrobacterium tumefaciens so as to establish a new transient expression system. Then the new transient expression system was applied to analyze maize In5-2 promoter functional area to clear the position of cis-acting elements responsive to chloroacetanilide. In this experiment onion(Allium cepa) epidermal cells were transformed with beta-glucuronidase(GUS) gene via Agrobacterium tumefaciens. The effects of different concentration of acetosyringone (As), infection period, concentration of Agrobacterium tumefaciens and days of co-culture on the transient expression of GUS gene were analyzed to establish a new transient expression system. The result indicated that a higher transient express of GUS gene could be acquired from onion epidermal cells which were dipped into Agrobaterium suspension (OD600 adjusted to 0.8, concentration of As adjusted to 20 μmol/L) for 15 min, and co-cultured for 3 days. So a new transient expression system was established. Then different length of maize(Zea mays) In5-2 promoter deletions were cloned to vector pCAMBIA1301 and transformed into the new transient expression system to analyze its functional regions. Cis-acting responsive elements to chloroacetanilide could be located within the range of -220 to -143 bp upstream of ATG. The results indicated that the new transient expression system is a fast and effective way to analyze promoter function.
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Received: 02 March 2010
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