Information of the inserted DNA sequences

The transforming plasmid pDAB8264 contained a transfer DNA (T-DNA) region comprising three gene cassettes expressing the herbicide tolerance proteins 2mEPSPS derived from Zea mays,  AAD-12 from Delftia acidovorans, and PAT from Streptomyces viridochromogenes. Transformation of soybean was achieved through Agrobacterium tumefaciens-mediated transformation.

(1) Transcription of the 2mepsps coding sequence is directed by histone H4A748 promoter together with the 5′ untranslated region and an intron from the histone 3 gene from Arabidopsis thaliana. The coding region includes an optimized chloroplast transit peptide derived from Zea mays and Helianthus annuus RuBisCO sequences to direct the expressed protein to the chloroplast. Transcription is terminated by the 3′ untranslated region of the histone H4A748 gene from Arabidopsis thaliana. A matrix attachment region from the Nicotiana tabacum rb7 gene is also present to increase gene expression.  Expression of this cassette results in production of the 2mEPSPS protein containing two amino acid substitutions that confer reduced sensitivity to glyphosate, thereby allowing continued synthesis of aromatic amino acids through the shikimate pathway in the presence of the herbicide.

(2) Transcription of the aad-12 coding sequence is directed by the promoter together with the 5′ untranslated region and intron from the polyubiquitin 10 gene from Arabidopsis thaliana. Transcription is terminated by the  3' untranslated region (3' UTR) of open reading frame 23 (ORF23) of plasmid pTi15955 from Agrobacterium tumefaciens. Expression of this cassette results in production of the AAD-12 protein, an alpha-ketoglutarate-dependent dioxygenase that metabolically inactivates herbicides of the aryloxyalkanoate family, including 2,4-dichlorophenoxyacetic acid (2,4-D).
 

(3) Transcription of the pat coding sequence is directed by the promoter together with the 5′ untranslated region from the Cassava Vein Mosaic virus. Transcription is terminated by the 3' untranslated region (3' UTR) of the open reading frame 1 (ORF1) of plasmid pTi15955 from Agrobacterium tumefaciens. Expression of this cassette results in production of the PAT protein, a phosphinothricin acetyltransferase that inactivates glufosinate through acetylation.