Transgenic crop plants expressing synthetic cry9Aa gene are protected against insect damage

A synthetic gene sequence of cry9Aa was made to achieve high expression levels in a plant cell. Tobacco, potato, cauliflower and turnip rape plants were transformed with this synthetic gene driven by the double 35S promoter using Agrobacterium tumefaciens LBA4404. The presence and expression of the synthetic cry9Aa gene was evaluated in Southern, Northern and Western analysis and with insect bioassays. The expression of the gene in tobacco plants reached a level of 5 pg of mRNA per 1 μg of total RNA and 0.3% of soluble protein or 1.4 μg of Cry9Aa protein per 1 g of leaf material. The expression level in the other species was three to ten times lower. Tobacco plants were also transformed with a truncated native cry9Aa gene construct and with a translational fusion construct of the truncated native cry9Aa and the uidA (GUS) gene sequence. The constructs were transformed in tobacco plants under the control of the same promoter as the synthetic cry9Aa. The expression level of the native cry9Aa gene constructs ranged from 0.03 to 1 pg of cry9Aa mRNA per 1 μg of total RNA. The protein was undetectable in Western analysis. In comparison to the native constructs the expression level of the synthetic cry9Aa gene was five to ten times higher at the mRNA level and at least 50 times higher at the translational level. Bioassays against Plutella xylostella performed with transgenic cauliflower showed high insecticidal activity of the plants expressing the synthetic cry9Aa gene.