Sugarcane bacilliform viral (scbv) enhancer and its use in plant functional genomics

Abstract

Identification of new enhancer sequence has significant utility in the plant functional genomics. The sugarcane bacilliform badnavirus (SCBV) transcriptional enhancer has been identified. This enhancer can be used to increase the rate of transcription from gene promoters and in activation tagging experiments. A ten-fold increase in transcription was observed when a 4× array of the SCBV enhancer was placed upstream of a truncated form of the maize alcohol dehydrogenase minimal promoter. Methods of using the SCBV transcriptional enhancer are described, as are chimeric transcription regulatory regions, constructs, cells, tissues, and organisms that comprise one or more copies of the enhancer.

We claim: 
     
         1 . A chimeric transcription regulatory region comprising:
 one or more copies of the sugarcane bacilliform viral (SCBV) enhancer element shown in position 337 to position 618 of SEQ ID NO: 1, or a homolog thereof; and   operably linked thereto, a promoter comprising an RNA polymerase binding site and a mRNA initiation site,   wherein when a nucleotide sequence of interest is transcribed under regulatory control of the chimeric transcription regulatory region, the amount of transcription product is enhanced compared to the amount of transcription product obtained with the chimeric transcription regulatory region comprising the promoter and not comprising the SCBV enhancer sequence(s).   
     
     
         2 . The chimeric transcription regulatory region of  claim 1 , wherein the promoter is obtained from the upstream region of a plant virus gene, a bacterial gene, a fungal gene, a plant nuclear gene, a plant extra-nuclear gene, an invertebrate gene, or a vertebrate gene. 
     
     
         3 . A construct comprising the transcriptional initiation region of  claim 1  operably linked to a transcribable polynucleotide molecule operably linked to a 3′ transcription termination polynucleotide molecule. 
     
     
         4 . The construct of  claim 3 , wherein said transcribable polynucleotide molecule confers an agronomic trait to a plant in which it is expressed. 
     
     
         5 . A transgenic plant stably transformed with the construct of  claim 3 . 
     
     
         6 . The transgenic plant of  claim 5 , wherein the transcribable polynucleotide molecule confers an agronomic trait to a plant in which it is expressed. 
     
     
         7 . A seed of the transgenic plant of  claim 5 , wherein the seed comprises the construct. 
     
     
         8 . The transgenic plant of  claim 5 , which plant is a maize plant 
     
     
         9 . A transgenic plant cell comprising the chimeric transcription regulatory region of  claim 1 . 
     
     
         10 . A method of producing a transgenic plant comprising transforming a plant cell or tissue with the construct of  claim 3 . 
     
     
         11 . The method of  claim 10 , wherein the transgenic plant is a dicotyledon. 
     
     
         12 . The method of  claim 10 , wherein the transgenic plant is a monocotyledon. 
     
     
         13 . A plant cell or tissue transformed with the construct of  claim 3 . 
     
     
         14 . The plant cell or tissue of  claim 13 , wherein the plant cell or tissue is from a dicotyledon. 
     
     
         15 . The plant cell or tissue of  claim 13 , wherein the plant cell or tissue is derived from a monocotyledon. 
     
     
         16 . A plant cell, fruit, leaf, root, shoot, flower, seed, cutting and other reproductive material useful in sexual or asexual propagation, progeny plants inclusive of F1 hybrids, male-sterile plants and all other plants and plant products derivable from the transgenic plant of  claim 5 . 
     
     
         17 . A maize plant cell, tissue or plant comprising one or more copies of the sugarcane bacilliform viral (SCBV) enhancer element shown in position 337 to position 618 of SEQ ID NO: 1, or a homolog thereof, in which the one or more copies of the SCBV enhancer element is inserted into a genome of the maize plant cell, tissue or plant at a random location. 
     
     
         18 . The maize plant cell, tissue or plant of  claim 17 , wherein the SCBV enhancer imparts enhanced transcription of a nucleotide sequence of interest which is under regulatory control of the SCBV enhancer as compared to transcription of the nucleotide sequence of interest in the absence of the SCBV enhancer.