A plant cell comprising: a DNA construct having as components, (a) a duplicated CaMV 35s enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; and (ii) a promoter comprising an RNA polymerase binding site and an mRNA initiation site; (b) a nucleotide sequence of interest for transcription to mRNA; and (c) a termination region wherein said components are operably joined.

A DNA construct having as components, (a) a transcription initiation region including (i) a tandemly duplicated CaMV 35S enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; (ii) a promoter comprising an RNA polymerase binding site and an mRNA initiation site; (b) a nucleotide sequence of interest for transcription to mRNA; and (c) a termination region wherein said components are operably joined.

A chimeric transcriptional initiation region comprising: as operably joined components (i) a tandemly duplicated CaMV 35S enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; and (ii) a promoter comprising an RNA polymerase binding site and an mRNA initiation site, wherein when a nucleotide sequence of interest is transcribed under the regulatory control of said chimeric transcriptional initiation region, the amount of transcription product is enhanced as compared to the amount of transcription obtained with a chimeric transcriptional initiation region comprising a single copy of said CaMV enhancer sequence and said promoter.

A differentiated plant comprising that comprise a DNA construct having as components (a) a tandemly duplicated CaMV 35S enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; and (b) a promoter comprising an RNA polymerase binding site and an mRNA initiation site; (c) a nucleotide sequence of interest; and (d) a termination region; wherein said components are operably joined.

Claim 1
A method for expressing a DNA sequence of interest comprising transforming a plant cell with a DNA construct comprising (a) a tandemly duplicated CaMV 35S enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; (b) a promoter comprising an RNA polymerase binding site and an mRNA initiation site; (c) a nucleotide sequence of interest; and (d) a termination region; wherein said components are operably joined so that said sequence of interest is transcribed.

A transcriptional initiation region comprising:
(a) a plant or viral enhancer domain comprising a plurality of units each including a sequence of the group comprising: GTGG, ATGG, GTAG, GTGA, GTGGA(T)A(T)A(T),GTGTGGA(T)A(T)A(T)G, and complementary sequence thereof, said enhancer domain further comprising at least one additional unit including a sequence selected from the said group; and
(b) a transcription initiation domain comprising an RNA polymerase binding site and an mRNA initiation site, under the enhancing control of said enhancer domain.

A DNA construct comprising: a transcription initiation region comprising:
(a) an enhancer domain comprising a plurality of units each including a sequence selected from the group comprising: GTGG, ATGG, GTAG, GTGA, GTGGA(T)A(T)A(T),GTGTGGA(T)A(T)A(T)G, and complementary sequence thereof;  and
(b) a transcription initiation domain comprising an RNA polymerase binding site and an mRNA initiation site under the enhancing control of said enhancer domain;
(c) a sequence of interest for transcription to mRNA; and
(d) a termination region.

A DNA sequence comprising a transcription initiation region capable of regulating transcription of a nucleotide sequence of interest in a plant cell and including, as operably joined components, (i) two to four copies in tandem of an enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; and (ii) a promoter comprising an RNA polymerase binding site and an mRNA initiation site.

A chimeric transcriptional initiation region comprising: as operably joined components, (i) two or more copies of a CaMV 35S enhancer sequence and (ii) a promoter comprising an RNA polymerase binding site and an mRNA initiation site, wherein when a nucleotide sequence of interest is transcribed under the regulatory control of said chimeric transcriptional initiation region, the amount of transcription product is enhanced as compared to the amount of transcription obtained with a chimeric transcriptional initiation region comprising a single copy of said CaMV enhancer sequence and a promoter.

A DNA construct having as components, (a) a transcription initiation region including (i) a tandemly duplicated CaMV 35S enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; (ii) a promoter comprising an RNA polymerase binding site and an mRNA initiation site; (b) a nucleotide sequence of interest for transcription to mRNA; and (c) a termination region wherein said components are operably joined.

A method for producing a plant cell capable of enhanced expression of a nucleotide sequence of interest, said method comprising: operably linking to form an expression cassette said nucleotide sequence of interest and a transcription initiation region capable of regulating transcription of a DNA sequence of interest in a plant cell, wherein said transcription initiation region comprises (i) two to four copies in tandem of an enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; and (ii) a promoter comprising an RNA polymerase binding site and an mRNA initiation site; and introducing said expresssion cassette into said plant cell to form a plant cell capable of enhanced expression of a said nucleotide sequence of interest.

A plant cell comprising: a DNA construct having components, (a) a transcription initiation region including (i) a tandemly duplicated CaMV 35S enhancer sequence comprising an AluI-EcoRV fragment of a CaMV 35S upstream region; (ii) a promoter comprising an RNA polymerase binding site and an mRNA initiation site; (b) a nucleotide sequence of interest for transcription to mRNA; and (c) a termination region wherein said components are operably joined.