Linked Life Data

11772640

Source:http://linkedlifedata.com/resource/pubmed/id/11772640

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pubmed-article:11772640pubmed:abstractTextThe 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.lld:pubmed
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pubmed-article:11772640pubmed:authorpubmed-author:LinzJohn EJElld:pubmed
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pubmed-article:11772640pubmed:dateRevised2010-9-14lld:pubmed
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pubmed-article:11772640pubmed:articleTitleChromosomal location plays a role in regulation of aflatoxin gene expression in Aspergillus parasiticus.lld:pubmed
pubmed-article:11772640pubmed:affiliationDepartment of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan 48824, USA.lld:pubmed
pubmed-article:11772640pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:11772640pubmed:publicationTypeResearch Support, U.S. Gov't, P.H.S.lld:pubmed
pubmed-article:11772640pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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