Corn event PV-ZMGT32(nk603) and compositions and methods for detection thereof

Abstract

The present invention provides a DNA construct that confers tolerance to transgenic corn plant. Also provided are assays for detecting the presence of the PV-ZMGT32(nk603) corn event based on the DNA sequence of the recombinant construct inserted into the corn genome and of genomic sequences flanking the insertion site.

1 . A DNA construct comprising: a first and a second expression cassette, wherein said first expression cassette in operable linkage comprises (i) a rice actin 1 promoter; (ii) a rice actin 1 intron; (iii) a chloroplast transit peptide encoding DNA molecule; (iv) a glyphosate tolerant EPSPS encoding DNA molecule; and (v) a transcriptional terminator DNA molecule; and said second expression cassette comprising in operable linkage (a) a CaMV 35S promoter; (b) a Hsp70 intron; (c) a chloroplast transit peptide encoding DNA molecule; (d) a glyphosate tolerant EPSPS encoding DNA molecule; and (e) a transcriptional terminator DNA molecule.  
     
     
         2 . A DNA construct of  claim 1 , wherein the glyphosate tolerant EPSPS encoding DNA molecule consists of an AGRTU.aroA:CP4 DNA molecule.  
     
     
         3 . A plant comprising the DNA construct of  claim 2 .  
     
     
         4 . A plant of  claim 3 , wherein said plant is a corn plant.  
     
     
         5 . A corn plant provided by ATCC seed deposit PTA-2478, progeny seeds and plants or parts thereof.  
     
     
         6 . A DNA molecule comprising a nucleotide sequence identified as SEQ ID NO:7 or SEQ ID NO:8.  
     
     
         7 . A pair of DNA molecules comprising: a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of sufficient length of contiguous nucleotides of SEQ ID NO:7 or its complement to function as DNA primers or probes diagnostic for DNA extracted from corn plant PV-ZMGT32(nk603) or progeny thereof.  
     
     
         8 . A pair of DNA molecules comprising: a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of sufficient length of contiguous nucleotides of SEQ ID NO:8 or its complement to function as DNA primers or probes diagnostic for DNA extracted from corn plant PV-ZMGT32(nk603) or progeny thereof.  
     
     
         9 . A method of detecting the presence of a DNA molecule in a corn plant provided by ATCC seed deposit PTA-2478 or progeny thereof, the method comprising: 
 (a) extracting a DNA sample from said corn plant provided by ATCC seed deposit PTA-2478 or progeny seeds and plants or parts thereof;    (b) contacting the DNA sample with a DNA primer pair comprising DNA primer molecules of sufficient length of contiguous nucleotides of SEQ ID NO:7 or its complement, or SEQ ID NO:8 or its complement;    (c) providing a nucleic acid amplification reaction condition;    (d) performing said nucleic acid amplification reaction, thereby producing a DNA amplicon molecule comprising the DNA molecules selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12; and    (e) detecting the DNA amplicon molecule.    
     
     
         10 . A corn plant when analyzed by a method of  claim 9  produced an amplicon comprising the DNA molecules selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12.  
     
     
         11 . In the method of  claim 9 , the DNA amplicon molecule comprising the DNA molecules selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12.  
     
     
         12 . A method of detecting the presence of a DNA molecule selected from the group consisting of SEQ ID NO:7 and SEQ ID NO:8 in a DNA sample, the method comprising: 
 (a) extracting a DNA sample from a corn plant;    (b) contacting the DNA sample with a DNA molecule identified as SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12, wherein said DNA molecule is a DNA probe that hybridizes under stringent hybridization conditions with the DNA molecule selected from the group consisting of SEQ ID NO:7 or SEQ ID NO:8, and does not hybridize under the stringent hybridization conditions with a DNA sample not containing the DNA molecule identified as SEQ ID NO:7 or SEQ ID NO:8;    (c) subjecting the sample and probe to stringent hybridization conditions; and    detecting hybridization of the probe to the DNA.    
     
     
         13 . A DNA molecule selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, and complements thereof.  
     
     
         14 . A method of breeding a glyphosate tolerant trait into corn plants comprising 
 a) extracting a DNA sample from progeny corn plants;    b) contacting the DNA sample with a marker nucleic acid molecule selected from the group consisting of SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, and complements thereof.    c) performing a marker assisted breeding method for the glyphosate tolerant trait, wherein the glyphosate tolerant trait is genetically linked to a complement of the marker nucleic acid molecule;    
     
     
         15 . A method of producing a corn plant that tolerates application of glyphosate herbicide comprising: 
 (a) transforming a corn cell with the DNA construct of  claim 1;     (b) selecting said corn cell for tolerance to application of glyphosate;    (c) growing said corn cell into a fertile corn plant;    
     
     
         16 . A DNA detection kit comprising: at least one DNA molecule of sufficient length of contiguous nucleotides homologous or complementary to SEQ ID NO:7 or SEQ ID NO:8 that functions as a DNA primer or probe specific for corn event PV-ZMGT32(nk603) and its progeny.