Genetic elements introduced from PV-ZMAP1043:
The expression of Agrobacterium tumefaciens 5-enolpyruvylshikimate-3-phosphate synthase (cp4 epsps) is under the control of the Cauliflower Mosaic Virus 35S enhanced promoter (P-e35S) and the A. tumefaciens nopaline synthase (nos) terminator. The transcript contains the Zea Mays heatshock protein 70 intron (I-Hsp70) for improved in planta expression and the Arabidopsis thaliana chloroplast transit peptide 2 for targeting the protein to the chloroplast. I-Hsp70 is not translated. The cp4 epsps coding sequence is the codon optimized coding sequence of the aroA gene from A. tumefaciens strain CP4 encoding CP4 EPSPS.

Southern blot analyses indicate that a single copy of the T-DNA was inserted at a single site in the maize genome and no plasmid vector backbone sequences were detected to have been integrated. DNA sequencing analyses indicated that the expected T-DNA sequences were integrated.

Genetic elements introduced from PV-ZMGT32:
The plant expression plasmid vector, PV-ZMGT32 contains two adjacent plant gene expression cassettes each containing a single copy of the cp4 epsps. In the first (5' end) expression cassette, the cp4 epsps gene is under the regulation of the Oryza sativa actin promoter (P-Ract1) and the rice actin intron (I-Ract1). The second cassette, which is fused to the 3' end of the first, consists of the cp4epsps gene regulated by P-e35S  and I-Hsp70. Both expression cassettes incorporate the 3'untranslated region of the nos for signal polyadenylation.

The vector also contains the nptII gene encoding kanamycin resistance allowing selection of bacteria containing the plasmid, and an origin of replication (ori) necessary for replicating the plasmid in Escherichia coli. For the transformation through particle bombardment, a fragment of the vector (obtained after digestion with the restriction enzyme MluI) was used which contains only the two cp4 epsps gene expression cassettes. Therefore, the nptII gene and the origin of replication were not inserted into NK603.

Corn NK603 contains one insertion site containing a single copy of the linear DNA of PV-ZMGT32 used for transformation. Both cp4 epsps gene cassettes within the single insert which are intact.

Note:
* P-e35S is a 0.61Kb long sequence containing the promoter and leader for the cauliflower mosaic virus (CaMV) 35S RNA containing the duplicated enhancer region. This modification was made to enhance the activity of this promoter in plants.)
** For more information, please refer to the parental LMO records.