Line GTSB77 was produced using an Agrobacterium-meditated transformation protocol to transform B. vulgaris subsp. vulgaris A1012.  This technique is well studied (Klee and Rogers, 1989; and Zambryski, 1988).  Transformed material was treated with chemicals and checked to ensure that Agrobacterium was not present.<br>The EPSPS gene codes for an enzyme that catalyzes the conversion of shikimate-3-phosphate and phosphoenolpyruvate into 5-enolpyruvlshikmate-3-phosphate, an intermediate in the production of aromatic amino acids.  This enzyme is present in many plants and microorganisms.  The enzyme used here is insensitive to the herbicide glyphosate unlike the EPSPS that is found in sugar beets.  The original sequence from Agrobacterium was modified to create a synthetic gene that allows higher expression of bacterial gene sequences in plants. The EPSPS present in sugar beet line GTSB77 is functionally and structurally similar to EPSPS protein present in food and feeds derived from plant and microbial sources.  Comparing the deduced amino acid sequences of  EPSPS from Agrobacterium sp. with EPSPS from soybean, corn, petunia, E. coli, Bacillus subtilis, and Saccharomyces cerevisiae (Baker's Yeast) yield amino acid similarities in the range of 48.5 to 59.3% and identities in the range of 23.2 to 41.4% (Padgette  et al. 1996).  The amino acid homologies observed between CP4 EPSPS and EPSPSs present in plants and bacteria is comparable to the homology between the EPSPSs from, for instance, soybean and B. subtilis (55.6 similar, 30.1% identical).  Thus, while there is some sequence diversity in EPSPSs      typically found in food and feed, the sequence variation between the EPSPS in GTSB77 and EPSPSs found in currently available food or feed is of the same magnitude (Padgette  et al. 1996).  <br>Data provided by Novartis/Monsanto demonstrate that line GTSB77 contains the following sequences:  (1) the 5-enolpyruvalshikimate-3-phosphate synthase gene from A. tumefaciens whose transcription is directed by the figwort mosaic virus promoter (Gownda et al., 1989), a chloroplast transit peptide from Arabidopsis thialiana, and whose termination/polyadenylation sequences were derived from the ribulose biphosphate carboxylase gene from pea (Coruzzi et al, 1984);  (2)  the beta-glucuronidase gene from E. coli whose transcription is directed by the enhanced CaMV promoter (Kay et al., 1985) and the same pea terminator sequences used for the EPSPS gene; and (3) a truncated version of the glyphosate oxidoreductase gene from O. anthropi whose transcription is directed by the figwort mosaic promoter and chloroplast transit peptide sequences that are described above.  This gene was inserted in an open reading frame of an uncharacterized plant gene resulting in the synthesis of a new protein that has a molecular weight of 35,000.  Also, inserted is the right border sequence from A. tumefaciens.  <br>Novartis/Monsanto presented data that the following sequences that were present in the original plasmid in Agrobacterium were not present in GTSB77:  the aad gene that confers resistance to the antibiotic streptomycin, neomycin phosphotransferase that confers resistance to kanamycin/neomycin, and oriV/ori 322  both involved in plasmid replication in Agrobacterium.