Blackcurrant promoters and genes

Abstract

Promoters and a process for isolating a promoter capable of driving fruit-specific expression of DNA sequences in transgenic blackcurrant and other non-climacteric fruit.

1 . A process for isolating a promoter capable of driving fruit-specific expression of DNA sequence in transgenic blackcurrent and other non-climacteric fruit comprising: 
 a) isolating mRNA from ripening blackcurrant fruit    b) preparing a cDNA library from the isolated mRNA    c) differentially screening the library from b) to identify genes expressed during the ripening period    d) screening a genomic library with probes prepared from cDNA identified according to step c) to isolate the corresponding gene and its promoter region.    
     
     
         2 . A promoter capable of driving fruit-specific expression of DNA sequences in transgenic blackurrant and other non-climacteric fruit obtainable by the process of  claim 1 .  
     
     
         3 . A promoter according to  claim 2  which comprises the sequence of nucleic acid bases in FIG. 9 or IDSEQ 11 (the RIB1 gene promoter) or IDSEQ 14 (the RIB 7 gene promoter.  
     
     
         4 . Promoter DNA sequences which hybridise to the DNA of  claim 3  under conditions of high stringency.  
     
     
         5 . cDNA for genes which exhibit differential expression in fruit during the ripening period of fruit development selected from pRIB1 (IDSEQ 1), pRIB3 (IDSEQ 3), pRIB5 (IDSEQ 5), pRIB6 (IDSEQ 7) and pRIB7 (IDSEQ 9).  
     
     
         6 . DNA encoding the RIB1 or RIB7 gene.  
     
     
         7 . A vector comprising the DNA as claimed in any one of  claims 2  to  6 .  
     
     
         8 . Use of a promoter according to  claim 2 , 3  or  4  to control the expression of one or more genes in climacteric or non-climacteric fruit.  
     
     
         9 . Use according to  claim 8  wherein the non-climacteric fruit is blackcurrant.  
     
     
         10 . Use of a promoter according to  claim 2 , 3  or  4  in the transformation of plant cells.  
     
     
         11 . Plant cells and plants transformed using a promoter according to claims  2 , 3  or  4  or a vector according to  claim 7 .  
     
     
         12 . Plants comprising cells according to  claim 11  and descendants thereof.  
     
     
         13 . Plants and seeds according to  claim 12  which are blackcurrants and products prepared therefrom.  
     
     
         14 . A process according to  claim 1  wherein the method for extracting nucleic acid from blackcurrant fruit comprises homogenising by pulping blackcurrant fruit in a buffer containing insoluble polyvinvylpolypyrrolidone.  
     
     
         15 . Proteins encoded by the DNA sequences of claims  5  or  6 .