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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
2002-1-4
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pubmed:abstractText |
The nor-1 gene in the filamentous fungus Aspergillus parasiticus encodes a ketoreductase involved in aflatoxin biosynthesis. To study environmental influences on nor-1 expression, we generated plasmid pAPGUSNNB containing a nor-1 promoter-beta-glucuronidase (GUS) (encoded by uidA) reporter fusion with niaD (encodes nitrate reductase) as a selectable marker. niaD transformants of A. parasiticus strain NR-1 (niaD) carried pAPGUSNNB integrated predominantly at the nor-1 or niaD locus. Expression of the native nor-1 and nor-1::GUS reporter was compared in transformants grown under aflatoxin-inducing conditions by Northern and Western analyses and by qualitative and quantitative GUS activity assays. The timing and level of nor-1 promoter function with pAPGUSNNB integrated at nor-1 was similar to that observed for the native nor-1 gene. In contrast, nor-1 promoter activity in pAPGUSNNB and a second nor-1::GUS reporter construct, pBNG3.0, was not detectable when integration occurred at niaD. Because niaD-dependent regulation could account for the absence of expression at niaD, a third chromosomal location was analyzed using pAPGUSNP, which contained nor-1::GUS plus pyrG (encodes OMP decarboxylase) as a selectable marker. GUS expression was detectable only when pAPGUSNP integrated at nor-1 and was not detectable at pyrG, even under growth conditions that required pyrG expression. nor-1::GUS is regulated similarly to the native nor-1 gene when it is integrated at its homologous site within the aflatoxin gene cluster but is not expressed at native nor-1 levels at two locations outside of the aflatoxin gene cluster. We conclude that the GUS reporter system can be used effectively to measure nor-1 promoter activity and that nor-1 is subject to position-dependent regulation in the A. parasiticus chromosome.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-10075929,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-10584035,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-15012504,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-1910523,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-2176447,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-2277647,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-2525275,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-2689886,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-3039345,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-3549463,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-3550422,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-3606105,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-6230518,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-7034956,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-7546572,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-7618880,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-7773383,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-8056329,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-8517755,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-9055421,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-942051,
http://linkedlifedata.com/resource/pubmed/commentcorrection/11772640-9532734
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0099-2240
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
68
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
306-15
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pubmed:dateRevised |
2010-9-14
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pubmed:meshHeading |
pubmed-meshheading:11772640-Aflatoxins,
pubmed-meshheading:11772640-Alcohol Oxidoreductases,
pubmed-meshheading:11772640-Aspergillus,
pubmed-meshheading:11772640-Chromosomes, Fungal,
pubmed-meshheading:11772640-Fungal Proteins,
pubmed-meshheading:11772640-Gene Expression Regulation, Fungal,
pubmed-meshheading:11772640-Genes, Reporter,
pubmed-meshheading:11772640-Glucuronidase,
pubmed-meshheading:11772640-Nitrate Reductase,
pubmed-meshheading:11772640-Nitrate Reductases,
pubmed-meshheading:11772640-Plasmids,
pubmed-meshheading:11772640-Polymerase Chain Reaction,
pubmed-meshheading:11772640-Promoter Regions, Genetic,
pubmed-meshheading:11772640-Recombinant Fusion Proteins,
pubmed-meshheading:11772640-Restriction Mapping,
pubmed-meshheading:11772640-Transformation, Genetic
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pubmed:year |
2002
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pubmed:articleTitle |
Chromosomal location plays a role in regulation of aflatoxin gene expression in Aspergillus parasiticus.
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pubmed:affiliation |
Department of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan 48824, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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