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Volumn 277, Issue 5326, 1997, Pages 696-699

Analysis of a chemical plant defense mechanism in grasses

Author keywords

[No Author keywords available]

Indexed keywords

HYDROXAMIC ACID DERIVATIVE;

EID: 15444344334     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5326.696     Document Type: Article
Times cited : (556)

References (32)
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    • Individual plants were analyzed by PCR amplification of DNA restriction fragments ligated to an adapter, similar to the amplified fragment length polymorphism (AFLP) protocol [P. Vos et al., Nucleic Acids Res. 23, 4407 (1995)]; however, a Mu-specific biotinylated primer [AGAGAAGCCAACGCCA(A/T)C-GCCTCCATT] was used to isolate Mu-sequences.
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    • note
    • The maize inbred line C31A was the source for all wild-type cDNA and genomic clones. Cloning was performed as described (6).
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    • Isolation of indole-3-glycerol phosphate and fluorimetric enzymatic assay were done as described [T. E. Creighton, Eur. J. Biochem. 13, 1 (1970)]. A reaction volume of 1 ml containing 4 μg of BX1 protein, 50 μg of glyceraldehyde phosphate dehydrogenase, and 0.5 mM nicotinamide adenine dinucleotide (oxidized form) was incubated at 22°C for 2 min. Indole-3-glycerol phosphate concentration varied from 0 to 50 μM. The identity of the product indole was proven by Ehrlich's reagent [W. K. Lim et al., J. Bacteriol. 173, 1886 (1991)]. BX1 protein was expressed in E. coli by inserting the cDNA into a modified pET3a vector [A. H. Rosenberg et al., Gene 56, 125 (1987)] and was purified to homogeneity by means of a six-nucleotide oligomer COOH-terminal histidine tag (Qiagen Ni-NTA purification system).
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    • Isolation of indole-3-glycerol phosphate and fluorimetric enzymatic assay were done as described [T. E. Creighton, Eur. J. Biochem. 13, 1 (1970)]. A reaction volume of 1 ml containing 4 μg of BX1 protein, 50 μg of glyceraldehyde phosphate dehydrogenase, and 0.5 mM nicotinamide adenine dinucleotide (oxidized form) was incubated at 22°C for 2 min. Indole-3-glycerol phosphate concentration varied from 0 to 50 μM. The identity of the product indole was proven by Ehrlich's reagent [W. K. Lim et al., J. Bacteriol. 173, 1886 (1991)]. BX1 protein was expressed in E. coli by inserting the cDNA into a modified pET3a vector [A. H. Rosenberg et al., Gene 56, 125 (1987)] and was purified to homogeneity by means of a six-nucleotide oligomer COOH-terminal histidine tag (Qiagen Ni-NTA purification system).
    • (1991) J. Bacteriol. , vol.173 , pp. 1886
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    • Isolation of indole-3-glycerol phosphate and fluorimetric enzymatic assay were done as described [T. E. Creighton, Eur. J. Biochem. 13, 1 (1970)]. A reaction volume of 1 ml containing 4 μg of BX1 protein, 50 μg of glyceraldehyde phosphate dehydrogenase, and 0.5 mM nicotinamide adenine dinucleotide (oxidized form) was incubated at 22°C for 2 min. Indole-3-glycerol phosphate concentration varied from 0 to 50 μM. The identity of the product indole was proven by Ehrlich's reagent [W. K. Lim et al., J. Bacteriol. 173, 1886 (1991)]. BX1 protein was expressed in E. coli by inserting the cDNA into a modified pET3a vector [A. H. Rosenberg et al., Gene 56, 125 (1987)] and was purified to homogeneity by means of a six-nucleotide oligomer COOH-terminal histidine tag (Qiagen Ni-NTA purification system).
    • (1987) Gene , vol.56 , pp. 125
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    • 0029167503 scopus 로고
    • 1 maize plants mutagenized by means of a Robertson's Mutator element was screened for Mu-containing alleles of the P-450 genes by a reverse genetics-based technology [R. J. Bensen et al., Plant Cell 7, 75 (1995)]. PCR amplifications were done as described [M. Mena et al., Science 274, 1537 (1996)].
    • (1995) Plant Cell , vol.7 , pp. 75
    • Bensen, R.J.1
  • 19
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    • 1 maize plants mutagenized by means of a Robertson's Mutator element was screened for Mu-containing alleles of the P-450 genes by a reverse genetics-based technology [R. J. Bensen et al., Plant Cell 7, 75 (1995)]. PCR amplifications were done as described [M. Mena et al., Science 274, 1537 (1996)].
    • (1996) Science , vol.274 , pp. 1537
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    • G. Truan et al., Gene 125, 49 (1993).
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    • P. Urban et al., Eur. J. Biochem. 222, 843 (1994). Full-size P-450 cDNAs were inserted in the expression vector, beginning with the AUG translation start codon. This was accomplished by PCR amplification of the relevant cDNA sequences.
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    • Urban, P.1
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    • note
    • 1H NMR (Table 1).
  • 31
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    • note
    • We used gradient-enhanced heteronuclear multiple-quantum coherence, heteronuclear multiple-bond correlation, and double-quantum filtered correlation spectroscopy techniques (M. Frey et al., data not shown).
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    • note
    • -13C]indole; M. H. Zenk, T. M. Kutchan, and K. Kirschner for discussions and suggestions; J. Winkler and H. Krause for GC-MS analysis; and P. Urban and D. Pompon for the yeast expression system. Supported by Deutsche Forschungsgemeinschaft grant SFB 369 and by Fonds der Chemischen Industrie.


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