Nucleotides encoding stop codons in multiple reading frames and methods of use

Abstract

Compositions having polynucleotides encoding multiple translational stop signals in more than one reading frame are provided. The compositions include isolated polynucleotides, expression cassettes, and vectors, as well as host cells, prokaryotic organisms, and eukaryotic organisms comprising the polynucleotide(s). Methods include using the polynucleotides to stop translation of an mRNA into a protein, to produce a transformed cell and/or organism comprising the polynucleotide, and to identify transformed cells or organisms of a specific lineage.

1 . An isolated polynucleotide encoding stop codons in all six reading frames, wherein the polynucleotide is from 12 to 450 nucleotides. 
     
     
         2 . The isolated polynucleotide of  claim 1 , wherein the polynucleotide is substantially identical to a polynucleotide of SEQ ID NO: 1-7. 
     
     
         3 . The isolated polynucleotide of  claim 1 , wherein the polynucleotide is selected from the group consisting of SEQ ID NO: 1-7. 
     
     
         4 . A recombinant T-DNA expression cassette comprising a polynucleotide encoding stop codons in all six reading frames, wherein the polynucleotide is from 12 to 450 nucleotides. 
     
     
         5 . (canceled) 
     
     
         6 . A non-human host cell comprising the polynucleotide of any one of  claims 1 - 3 . 
     
     
         7 . The host cell of  claim 6  wherein the host cell is a plant cell. 
     
     
         8 . The host cell of  claim 6  wherein the host cell is an  Agrobacterium    
     
     
         9 . A transformed plant comprising the polynucleotide of any one of  claims 1 - 3 . 
     
     
         10 . A transformed seed comprising the polynucleotide of any one of  claims 1 - 3 . 
     
     
         11 . The plant of  claim 9  wherein the plant is a monocot or a dicot. 
     
     
         12 . The plant of  claim 11  wherein the plant is maize. 
     
     
         13 . A method to produce a transformed host cell comprising:
 a) introducing a recombinant T-DNA expression cassette a polynucleotide encoding stop codons in all six reading frames, wherein the polynucleotide is from 12 to 450 nucleotides into the host cell; and,   b) selecting a transformed host cell having the polynucleotide inserted into its genome.   
     
     
         14 . The method of  claim 13 , wherein insertion of the polynucleotide into the host genome truncates translation of any unintended messenger RNA transcript. 
     
     
         15 . The method of claim, wherein insertion of the polynucleotide into the host genome stimulates degradation of any unintended messenger RNA transcript. 
     
     
         16 . The method of  claim 13 , wherein insertion of the polynucleotide into the host genome provides a means to identify any progeny derived from the transformed host cell. 
     
     
         17 . The method of  claim 16 , wherein the progeny comprise proprietary germplasm. 
     
     
         18 . The method of any one of  claims 13 - 17  wherein the host cell is selected from the group consisting of a plant cell, a bacterial cell, or a mammalian cell. 
     
     
         19 . The method of any one of  claims 13 - 18 , wherein the host cell is a plant cell. 
     
     
         20 . The method of any one of  claims 13 - 19 , wherein the host cell is a maize cell. 
     
     
         21 . The recombinant T-DNA expression cassette of  claim 4 , wherein the polynucleotide is substantially identical to a polynucleotide selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7. 
     
     
         22 . The recombinant T-DNA expression cassette of  claim 4 , wherein the polynucleotide is a polynucleotide of selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7. 
     
     
         23 . A method to produce a transformed host cell of  claim 13 , wherein the polynucleotide is substantially identical to a polynucleotide selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7. 
     
     
         24 . A method to produce a transformed host cell of  claim 13 , wherein the polynucleotide is a polynucleotide of selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7.