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note
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PYP-phytochrome was amplified by PCR with oligonucleotides GTSATCGGNAAGAACTTCTTY and ACSCGYTTSACGAANANCCA. The entire gene was cloned from a genomic library, sequenced, and deposited in GenBank database under accession number AF064527.
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24
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0031127716
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2,0.1 mM EDTA, and 50% glycerol p-Hydroxycinnamic acid was attached as described [S. Devanathan et al., Arch. Biochem. Biophys. 340, 83 (1997)].
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0344335032
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note
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32P-Labeled protein bands were quantified by Phosphorlmager analysis (Molecular Dynamics, Sunnyvale, CA).
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30
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0344767069
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note
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-1, so the maximum ratio of absorption of Ppr at 434 nm to that at 280 nm is 0.442. The ratio of absorption of reconstituted Ppr at 434 nm to that at 280 nm was 0.29, giving a ∼60% efficiency of attachment (7).
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31
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0345629578
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note
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ppr amino acid residues 114 to 750 were replaced with a spectinomycin resistance gene in the suicide vector pGmLacZ. After delivery by conjugation (27), double recombinants were selected and confirmed by PCR analysis.
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34
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0344335030
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GenBank Accession number AF121274
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GenBank Accession number AF121274.
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35
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0345629575
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note
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The K. centenum chs promoter was amplified by PCR with primers TACGTACCGATGAACACCCAGCCCAC and TAGCATGCCGTGAAAACGGGGGAGAG. The PCR product was cloned into the lacZ reporter plasmid pZJD11 (21) and maintained with antibiotics.
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36
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0345197521
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note
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R. centenum was cultured in peptone-yeast extractsoytone (12) and illuminated with infrared light (>700 nm). White light was provided with a 400-W pressure sodium Lumalux lamp (LU400, Osram Sylvania, Danvers, MA) and blue light (400 to 450 nm) by passing the white light through filters. Cells were assayed for β-Gal activity as described (21).
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0344767065
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Supplemental Web material is available at www. sciencemag.org/feature/data/1039035.shl.
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0344335027
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note
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We thank T. Meyer and R. Hangarter for comments. Supported by NIH grant GM 40941 (C.E.B.) and a NSF postdoctoral fellowship (B.G.R.).
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