Regulation of translation of heterologously expressed genes

Abstract

The present invention pertains to a method of expressing a protein of interest, preferably a heterologous protein, in preferably a plant. In a preferred embodiment said plant is a doubled haploid homozygous transgenic Nicotiana tabacum plant silenced for Ntp303. Furthermore, the invention relates to said plant with or without nucleic acid constructs according to the invention. Propagation, harvest and tissue material of said transgenic Nicotiana tabacum plant is also a part of the invention.

1 . A nucleic acid construct comprising a first nucleotide sequence that has at least 90% nucleotide sequence identity with the nucleotide sequence of SEQ ID No. 1, operably linked to a second nucleotide sequence encoding a protein. 
     
     
         2 . The nucleic acid construct of  claim 1 , wherein the nucleotide sequence encoding the protein is further operably linked to a promotor. 
     
     
         3 . The nucleic acid construct of  claim 1 , wherein the first nucleotide sequence comprises the sequence of SEQ ID NO:1. 
     
     
         4 . A recombinant animal, mammalian, plant, fungal or bacterial host cell containing one or more copies of the nucleic acid construct as defined by  claim 1 . 
     
     
         5 . Use of the first nucleotide sequence as defined in  claim 1  for expressing a protein. 
     
     
         6 . The use of  claim 5 , wherein the first nucleotide sequence is used in a plant, fungal, bacterial, animal or mammalian cell. 
     
     
         7 . The use of  claim 6 , wherein the first nucleotide sequence is used in a cell-free translation system using RNAs derived from the nucleic acid construct as defined in  claim 1 . 
     
     
         8 . A method for expressing a polypeptide of interest in a cell, wherein the cell is a plant, fungal, or mammalian cell, comprising the steps of:
 a. providing a nucleic acid construct comprising:
 i. a first nucleotide sequence having at least 90% identity to SEQ ID NO: 1 operably linked to 
 ii. a second nucleotide sequence encoding a polypeptide of interest, and further operably linked to 
 iii. a promoter; and 
   b. transforming the cell with said nucleic acid construct to obtain a transformed cell; and   c. expressing the polypeptide in said transformed cell.   
     
     
         9 . The method of  claim 8 , wherein the first nucleotide sequence has at least 95% homology to SEQ ID NO: 1. 
     
     
         10 . The method of  claim 8 , wherein the polypeptide of interest is heterologous to said cell. 
     
     
         11 . The method of  claim 8 , wherein the polypeptide is expressed in a cell of a doubled haploid homozygous transgenic Nicotiana tabacum plant silenced for ntp303. 
     
     
         12 . The method of  claim 10 , wherein the polypeptide is expressed in a cell of a doubled haploid homozygous transgenic Nicotiana tabacum plant silenced for ntp303. 
     
     
         13 . The method of  claim 8 , wherein the polypeptide is expressed in the pollen or seed of a plant that comprises said plant cell. 
     
     
         14 . The method of  claim 10 , wherein the polypeptide is expressed in the pollen or seed of a plant that comprises said plant cell. 
     
     
         15 . The method of  claim 8 , wherein the promoter of (a)(iii) is heterologous to said second nucleotide sequence. 
     
     
         16 . The method of  claim 8 , wherein the first nucleotide sequence comprises a sequence forming an HI and HII structure. 
     
     
         17 . The method of  claim 8 , wherein the first nucleotide sequence is DNA. 
     
     
         18 . The method of  claim 8 , wherein the first nucleotide sequence is RNA. 
     
     
         19 . The method of  claim 8 , further comprising step (d) recovering said polypeptide. 
     
     
         20 . The method of  claim 8 , wherein the cell is a fungal, plant or mammalian cell. 
     
     
         21 . The method of  claim 8 , wherein the cell is a plant or mammalian cell.