Gene Expression or Activity Enhancing Elements

Abstract

The present invention relates to transgenic nucleic acids, expression cassettes, vectors, plant cells, plant organs and plants. The invention also relates to methods for increasing expression or activity of a target gene, particularly in a plant cell or plant organ, and also to uses of recombinant nucleic acids and expression cassettes to increase expression or activity of a target gene or for manufacturing of a vector, plant cell, plant organ or plant. Incidentally, the invention relates to enhancers for achieving increased expression or activity of a target gene, particularly in a plant cell or plant organ, when operably linked to a promoter functional in such plant cell, plant organ or plant. The invention is described herein with reference to the technical field of production of polyunsaturated fatty acids (PUFAs), without being limited to this technical field. For the production of desired molecules in plant cells, e.g. PUFAs, it is frequently required to express a target gene heterologous to the plant cell, or to overexpress a target gene naturally found in said plant cell.

We claim: 
     
         1 . A recombinant nucleic acid, comprising a target gene and an untranslated region adjacent to the target gene, wherein the untranslated region comprises an enhancer of at least 18 consecutive nucleotides, wherein at least 14 nucleotides are adenosine or cytidine. 
     
     
         2 . A recombinant nucleic acid of  claim 1 , wherein the enhancer comprises any sequence according to SEQ ID NOS. 84, 85, 86, 87, 88 or 89. 
     
     
         3 . A recombinant nucleic acid of  claim 1 , wherein the enhancer comprises or consists of
 i) 18 consecutive nucleotides, of which at least 15 nucleotides are adenosine or cytidine, or   ii) 21 consecutive nucleotides, of which at least 15 nucleotides are adenosine or cytidine, or   iii) 22 consecutive nucleotides, of which at least 16 nucleotides are adenosine or cytidine or   iv) 24 consecutive nucleotides, of which at least 18nucleotides are adenosine or cytidine, or   v) 36 consecutive nucleotides, of which at least 27 nucleotides are adenosine or cytidine or   vi) 57 consecutive nucleotides, of which at least 42 nucleotides are adenosine or cytidine, or   vii) 83 consecutive nucleotides, of which at least 62 nucleotides are adenosine or cytidine.   
     
     
         4 . A recombinant nucleic acid of  claim 1 , wherein the enhancer comprises any sequence according to
 a) any of SEQ ID NOS. 20 to 45, or any sequence according to   b) the last 18, 21, 22, 24, 36 or 57 nucleotides of any of SEQ ID NOS.46-83, 161-170, 221-230, 276-285 or 301-310, or   c) any of SEQ ID NOS. 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103 or 137, or   d) a sequence according to b) or c) with 1 additional base inserted therein.   
     
     
         5 . A recombinant nucleic acid, comprising a plant promoter and an untranslated region adjacent to the promoter, wherein the untranslated region comprises an enhancer of at least 18 consecutive nucleotides, wherein at least 14 nucleotides are adenosine or cytidine. 
     
     
         6 . A recombinant nucleic acid according to  claim 5 , wherein the enhancer comprises or consists of
 i) 18 consecutive nucleotides, of which at least 15 nucleotides are adenosine or cytidine, or   ii) 21 consecutive nucleotides, of which at least 15 nucleotides are adenosine or cytidine, or   iii) 22 consecutive nucleotides, of which at least 16 nucleotides are adenosine or cytidine, or   iv) 24 consecutive nucleotides, of which at least 18 nucleotides are adenosine or cytidine, or   v) 36 consecutive nucleotides, of which at least 27 nucleotides are adenosine or cytidine, or   vi) 57 consecutive nucleotides, of which at least 42 nucleotides are adenosine or cytidine, or   vii) 83 consecutive nucleotides, of which at least 62 nucleotides are adenosine or cytidine.   
     
     
         7 . A recombinant nucleic acid of  claim 5 , wherein the enhancer comprises any sequence according to
 a) any of SEQ ID NOS. 20 to 45, or any sequence according to   b) the last 18, 21, 22, 24, 36 or 57 nucleotides of any of SEQ ID NOS. 46 to 83, or   c) any of SEQ ID NOS. 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103 or 137, or   d) a sequence according to b) or c) with 1 additional base inserted therein.   
     
     
         8 . A recombinant nucleic acid of  claim 1 , wherein the enhancer comprises
 a) a CCAAT-Box comprising SEQ ID NO. 100, and/or   b) a Dof1/MNB1a binding site comprising SEQ ID NO. 102.   
     
     
         9 . A recombinant nucleic acid of  claim 1 , wherein the target gene is a fatty acid desaturase or elongase gene. 
     
     
         10 . An expression cassette, comprising a recombinant nucleic acid of  claim 1  and a plant promoter,
 wherein the promoter comprises
 a TATA-box, and 
 a CPRF factor binding site, and 
 a TCP class I transcription factor binding site, and 
 a bZIP protein G-Box binding factor 1 binding site. 
 
 
     
     
         11 . An expression cassette of  claim 10 , wherein the promoter further comprises one or more of the following sequences:
 a Ry motif,   a prolamin box,   a Cis-element as in GAPDH promoters conferring light inducibility,   a SBF-1 binding site,   a Sunflower homeodomain leucine-zipper protein Hahb-4 binding site.   
     
     
         12 . An expression cassette of  claim 10  , wherein the promoter comprises or consists of
 a) a nucleic acid according to any of SEQ ID NOS. 141, 142, 144, 145, 147, 148, 150, 151, 153, 154, 156, 157, 159 to 310, or 
 b) a nucleic acid having at least 70% sequence identity to any of the nucleic acid sequences according to a). 
 
     
     
         13 . An expression cassette of  claim 10 , comprising or consisting of the sequence according to SEQ ID NO. 1 or of a sequence having at least 70% sequence identity to the sequence of SEQ ID NO. 1. 
     
     
         14 . An expression cassette of  claim 10 , wherein the target gene does not consist of a sequence according to SEQ ID NO. 311. 
     
     
         15 . A vector comprising an expression cassette of  claim 10 . 
     
     
         16 . A plant, plant organ or plant cell comprising
 a recombinant nucleic acid of  claim 1 , operably linked to a promotor.   
     
     
         17 . Method of increasing expression or activity of a target gene, comprising the steps of
 i) providing, upstream of the target gene, an untranslated region and a plant promoter to obtain an expression cassette of  claim 9 , and   ii) introducing the expression cassette into a plant cell.   
     
     
         18 . A plant, plant organ or plant cell comprising
 an expression cassette comprising a recombinant nucleic acid comprising a target gene and an untranslated region adjacent to the target gene, wherein the untranslated region comprises an enhancer of at least 18 consecutive nucleotides, wherein at least 14 nucleotides are adenosine or cytidine   and a plant promoter, wherein the promoter comprises
 a TATA-box, and 
 a CPRF factor binding site, and 
 a TCP class I transcription factor binding site, and 
 a bZIP protein G-Box binding factor 1 binding site.