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Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway

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Summary:Aspergillus terreus is an efficient producer of different extracellular enzymes which are used in food industries. Thus, we are interested in developing a genetic transformation system based on the nitrate structural gene niaD. In order to develop this system the nitrate assimilation pathway in A. terreus was studied and spontaneous mutants defective in the genes required for nitrate assimilation were obtained on the basis of chlorate resistance. Of particular interest were niaD mutants which failed to grow on nitrate but grew as wild type on other sole nitrogen sources. Afterwards the transformation system was developed for mutants defective in nitrate reductase of A. terreus using pSTA10 vector containing niaD gene from A. niger. The transformation frequency obtained was c.a. 0.5 per ug DNA. The vector appeared to be mitotically stable and Southern hybridization analysis of transformants showed that transformation events occurred by integration into to the recipient genome. To assess the amount of product made by the transformants, nitrate reductase assays were carried out under inducing conditions. Experiments to improved transformation frequency and to characterize the integration are now in progress.
Main Authors:Ferraz, Elza
Other Authors:Lima, Nelson; Mota, M.
Year:1996
Country:Portugal
Document type:other
Access type:open access
Associated institution:Universidade do Minho
Language:English
Origin:RepositóriUM - Universidade do Minho
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author Ferraz, Elza
author2 Lima, Nelson
Mota, M.
author2_role author
author
author_facet Ferraz, Elza
Lima, Nelson
Mota, M.
author_role author
contributor_name_str_mv RepositóriUM - Universidade do Minho
country_str PT
creators_json_txt [{\"Person.name\":\"Ferraz, Elza\"},{\"Person.name\":\"Lima, Nelson\"},{\"Person.name\":\"Mota, M.\"}]
datacite.contributors.contributor.contributorName.fl_str_mv RepositóriUM - Universidade do Minho
datacite.creators.creator.creatorName.fl_str_mv Ferraz, Elza
Lima, Nelson
Mota, M.
datacite.date.Accepted.fl_str_mv 1996-01-01T00:00:00Z
datacite.date.available.fl_str_mv 2006-01-06T15:41:17Z
datacite.date.embargoed.fl_str_mv 2006-01-06T15:41:17Z
datacite.rights.fl_str_mv http://purl.org/coar/access_right/c_abf2
datacite.titles.title.fl_str_mv Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
dc.contributor.none.fl_str_mv RepositóriUM - Universidade do Minho
dc.creator.none.fl_str_mv Ferraz, Elza
Lima, Nelson
Mota, M.
dc.date.Accepted.fl_str_mv 1996-01-01T00:00:00Z
dc.date.available.fl_str_mv 2006-01-06T15:41:17Z
dc.date.embargoed.fl_str_mv 2006-01-06T15:41:17Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv https://hdl.handle.net/1822/3839
dc.language.none.fl_str_mv eng
dc.publisher.none.fl_str_mv University of Missouri. Fungal Genetics Stock Center
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.title.fl_str_mv Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_1843
description Aspergillus terreus is an efficient producer of different extracellular enzymes which are used in food industries. Thus, we are interested in developing a genetic transformation system based on the nitrate structural gene niaD. In order to develop this system the nitrate assimilation pathway in A. terreus was studied and spontaneous mutants defective in the genes required for nitrate assimilation were obtained on the basis of chlorate resistance. Of particular interest were niaD mutants which failed to grow on nitrate but grew as wild type on other sole nitrogen sources. Afterwards the transformation system was developed for mutants defective in nitrate reductase of A. terreus using pSTA10 vector containing niaD gene from A. niger. The transformation frequency obtained was c.a. 0.5 per ug DNA. The vector appeared to be mitotically stable and Southern hybridization analysis of transformants showed that transformation events occurred by integration into to the recipient genome. To assess the amount of product made by the transformants, nitrate reductase assays were carried out under inducing conditions. Experiments to improved transformation frequency and to characterize the integration are now in progress.
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fulltext.url.fl_str_mv https://repositorium.uminho.pt/bitstreams/1fcf73f7-6ff6-4448-a3ea-d81292e8b777/download
id rum_b862048a2a51dfd06e6a17b2a5d222ca
identifier.url.fl_str_mv https://hdl.handle.net/1822/3839
instacron_str repositorium
institution Universidade do Minho
instname_str Universidade do Minho
language eng
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organization_str_mv urn:organizationAcronym:repositorium
person_str_mv Ferraz, Elza
Lima, Nelson
Mota, M.
publishDate 1996
publisher.none.fl_str_mv University of Missouri. Fungal Genetics Stock Center
reponame_str RepositóriUM - Universidade do Minho
repository_id_str urn:repositoryAcronym:rum
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spelling engUniversity of Missouri. Fungal Genetics Stock CenterengAspergillus terreus is an efficient producer of different extracellular enzymes which are used in food industries. Thus, we are interested in developing a genetic transformation system based on the nitrate structural gene niaD. In order to develop this system the nitrate assimilation pathway in A. terreus was studied and spontaneous mutants defective in the genes required for nitrate assimilation were obtained on the basis of chlorate resistance. Of particular interest were niaD mutants which failed to grow on nitrate but grew as wild type on other sole nitrogen sources. Afterwards the transformation system was developed for mutants defective in nitrate reductase of A. terreus using pSTA10 vector containing niaD gene from A. niger. The transformation frequency obtained was c.a. 0.5 per ug DNA. The vector appeared to be mitotically stable and Southern hybridization analysis of transformants showed that transformation events occurred by integration into to the recipient genome. To assess the amount of product made by the transformants, nitrate reductase assays were carried out under inducing conditions. Experiments to improved transformation frequency and to characterize the integration are now in progress.application/pdfengMutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathwayFerraz, ElzaLima, NelsonMota, M.HostingInstitutionOrganizationalRepositóriUM - Universidade do Minhoe-mailmailto:repositorium@usdb.uminho.ptrepositorium@usdb.uminho.ptCITATIONFERRAZ, Elza; LIMA, Nelson; MOTA, Manuel - Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway. In European Congress on Fungal Genetics, 3, Munster, 1996 – “On-line abstracts from the European Congress on Fungal Genetics 3, held March 27-30, 1996, Munster, Germany” [Em linha]. Kansas City : University of Missouri. Fungal Genetics Stock Center, [c. 1996]. [Consult. 6 Jan. 2006]. Disponível em: http://www.fgsc.net/ecfg/ecfg3.html2006-01-06T15:41:17Z19961996-01-01T00:00:00ZHandlehttps://hdl.handle.net/1822/3839http://purl.org/coar/access_right/c_abf2open access12650 bytesother research producthttp://purl.org/coar/resource_type/c_1843otherhttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://repositorium.uminho.pt/bitstreams/1fcf73f7-6ff6-4448-a3ea-d81292e8b777/download
spellingShingle Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
Ferraz, Elza
status SINGLETON
title Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
title_full Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
title_fullStr Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
title_full_unstemmed Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
title_short Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
title_sort Mutagenesis and transformation of Aspergillus terreus based on the nitrate reductase pathway
url https://hdl.handle.net/1822/3839
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