Putrescine-n-methyltransferase promoter

EXAMPLE 1

Cloning of the NtPMT Promoter

[0040]Using the PMT cDNA sequence published by N. Hibi et al., Plant Cell 6, 723–725 (1994) we designed nested, divergent PCR primers for Inverse-PCR. Tobacco genomic DNA was cut with a variety of restriction endonucleases, ligated at low DNA concentrations (to promote circularization), and then used as a template for Inverse-PCR. The longest amplification product was obtained from genomic DNA that had been digested with NdeI. This fragment was cloned and sequenced. Sequence comparisons to the PMT cDNA showed sequence identity in the known regions, illustrating that the amplified DNA product was, indeed, the 5′ flanking region of the PMT gene. The DNA sequence of the NtPMT promoter is as follows:

[0041]GTATACCAAA AATCAATTCA ACCCCCAAAA CATAATACAACCAATGTTAA TGCAATATCT CTGCTGCTAT CACGAAGATAATTGTAGCTC ACGAAAGTAG GATACATTAT GTAGGTTACATCACATAGAG GTAATCTAAA GCTCCCAATA ATAAGATGTGTAATGTTGAT TATGTAGAAA TTTGCCAGGT TATTTAGAATAAACAAGAAG AGGAGAAAAA AAGTACAATT TAC

EXAMPLE 2

Transgenic Plants

[0043]The NtPMTI promoter set forth in Example 1 (SEQ ID NO: 1) was fused to the GUS gene in pBI101 and transformed into tobacco in accordance with standard techniques. See, e.g., U.S. Pat. No. 5,837,876 at Examples 4–6. Transgenic tobacco was stained for GUS activity using X-Gluc. Transgenic roots stained for GUS activity.

EXAMPLE 3

Deletion Mutants

[0044]Additional examples of PMT promoters of the present invention include deletion mutants of the promoter set forth in SEQ ID NO: 1 above. These include mutants with 5′ regions deleted, as set forth in SEQ ID NO: 2–6 below, as follows:

[0045]           AATCAATTCA ACCCCCAAAA CATAATACAA CCAATGTTAASEQ ID NO: 2TGCAATATCT CTGCTGCTAT CACGAAGATA ATTGTAGCTC ACGAAAGTAGGATACATTAT GTAGGTTACA TCACATAGAG GTAATCTAAA GCTCCCAATAATAAGATGTG TAATGTTGAT TATGTAGAAA TTTGCCAGGT TATTTAGAATAAACAAGAAG AGGAGAAAAA AAGTACAATT TACCTGAACT CTTGAATGTATCCTACAAAT AACCTAGACT TCATGGACGT CAGTTGTCAG TTTACTTTTGTTTTAATGGT ACATCATTTG TCAAATACTT TATTTGGATA AAAACAGTTTTGCCTAAGGA GTAAACAGAT CCGGAGTAAG AAAGCAGACG ATTAAAGCPATTTTTAAAAA AGGAGAGAGA AATTAATGAG CACACACATA TACTAGTGAAATTAGGGTAC TAATTTACTA ATAATTGCAC CGAGACAAAC TTATATTTTAGTTCCAAAAT GTCAGTCTAA CCCTGCACGT TGTAATAAAT TTTTAACTCTATTATATTAT ATCGAGTTGC GCCCTCCACT CCTCGGTGTC CAAATTGTATTTAAATGCAT AGATGTTTAA TGGGAGTGTA CAGCAAGCTT TCGGAAAATA

[0046]