A Method for the Re-isolation of an Autonomously Replicating Plasmid from Aspergillus Transformants
An autonomously replicating plasmid is expected to increase the frequency of Aspergillus transformation. To construct this type of plasmid, we developed a rapid method of re-isolating autonomously replicating plasmids from Aspergillus transformants. Transformants grown in MM medium under selective p...
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| Veröffentlicht in: | Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 1995, Vol.59 (6), p.1133-1134 |
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| container_end_page | 1134 |
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| container_issue | 6 |
| container_start_page | 1133 |
| container_title | Bioscience, biotechnology, and biochemistry |
| container_volume | 59 |
| creator | Ozeki, Kenji Hizume, Kazuhisa Kanda, Akihiro Hamachi, Masaaki Nunokawa, Yataro |
| description | An autonomously replicating plasmid is expected to increase the frequency of Aspergillus transformation. To construct this type of plasmid, we developed a rapid method of re-isolating autonomously replicating plasmids from Aspergillus transformants. Transformants grown in MM medium under selective pressure for 1-2 days were converted to protoplasts with a cell wall lytic enzyme (e.g. Yatalase). The protoplasts were lysed with phenol/chloroform followed by precipitation with ethanol. The total DNA was treated with RNaseA, re-precipitated with PEG, and then used to transform E. coli. These re-isolated plasmids were mainly the plasmid monomer. |
| doi_str_mv | 10.1271/bbb.59.1133 |
| format | Article |
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To construct this type of plasmid, we developed a rapid method of re-isolating autonomously replicating plasmids from Aspergillus transformants. Transformants grown in MM medium under selective pressure for 1-2 days were converted to protoplasts with a cell wall lytic enzyme (e.g. Yatalase). The protoplasts were lysed with phenol/chloroform followed by precipitation with ethanol. The total DNA was treated with RNaseA, re-precipitated with PEG, and then used to transform E. coli. These re-isolated plasmids were mainly the plasmid monomer.</description><identifier>ISSN: 0916-8451</identifier><identifier>EISSN: 1347-6947</identifier><identifier>DOI: 10.1271/bbb.59.1133</identifier><identifier>PMID: 7613000</identifier><language>eng</language><publisher>Tokyo: Taylor & Francis</publisher><subject>Aspergillus - genetics ; Biological and medical sciences ; Biotechnology ; DNA Replication ; DNA, Recombinant - biosynthesis ; Fundamental and applied biological sciences. 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To construct this type of plasmid, we developed a rapid method of re-isolating autonomously replicating plasmids from Aspergillus transformants. Transformants grown in MM medium under selective pressure for 1-2 days were converted to protoplasts with a cell wall lytic enzyme (e.g. Yatalase). The protoplasts were lysed with phenol/chloroform followed by precipitation with ethanol. The total DNA was treated with RNaseA, re-precipitated with PEG, and then used to transform E. coli. These re-isolated plasmids were mainly the plasmid monomer.</description><subject>Aspergillus - genetics</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>DNA Replication</subject><subject>DNA, Recombinant - biosynthesis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Methods. Procedures. 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Psychology</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Methods. Procedures. Technologies</topic><topic>Plasmids</topic><topic>Vectors (cloning, transfer, expression). Insertion sequences and transposons</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ozeki, Kenji</creatorcontrib><creatorcontrib>Hizume, Kazuhisa</creatorcontrib><creatorcontrib>Kanda, Akihiro</creatorcontrib><creatorcontrib>Hamachi, Masaaki</creatorcontrib><creatorcontrib>Nunokawa, Yataro</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Bioscience, biotechnology, and biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ozeki, Kenji</au><au>Hizume, Kazuhisa</au><au>Kanda, Akihiro</au><au>Hamachi, Masaaki</au><au>Nunokawa, Yataro</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Method for the Re-isolation of an Autonomously Replicating Plasmid from Aspergillus Transformants</atitle><jtitle>Bioscience, biotechnology, and biochemistry</jtitle><addtitle>Biosci Biotechnol Biochem</addtitle><date>1995</date><risdate>1995</risdate><volume>59</volume><issue>6</issue><spage>1133</spage><epage>1134</epage><pages>1133-1134</pages><issn>0916-8451</issn><eissn>1347-6947</eissn><abstract>An autonomously replicating plasmid is expected to increase the frequency of Aspergillus transformation. To construct this type of plasmid, we developed a rapid method of re-isolating autonomously replicating plasmids from Aspergillus transformants. Transformants grown in MM medium under selective pressure for 1-2 days were converted to protoplasts with a cell wall lytic enzyme (e.g. Yatalase). The protoplasts were lysed with phenol/chloroform followed by precipitation with ethanol. The total DNA was treated with RNaseA, re-precipitated with PEG, and then used to transform E. coli. These re-isolated plasmids were mainly the plasmid monomer.</abstract><cop>Tokyo</cop><pub>Taylor & Francis</pub><pmid>7613000</pmid><doi>10.1271/bbb.59.1133</doi><tpages>2</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0916-8451 |
| ispartof | Bioscience, biotechnology, and biochemistry, 1995, Vol.59 (6), p.1133-1134 |
| issn | 0916-8451 1347-6947 |
| language | eng |
| recordid | cdi_pubmed_primary_7613000 |
| source | J-STAGE Free; MEDLINE; Freely Accessible Japanese Titles; EZB-FREE-00999 freely available EZB journals; Free Full-Text Journals in Chemistry |
| subjects | Aspergillus - genetics Biological and medical sciences Biotechnology DNA Replication DNA, Recombinant - biosynthesis Fundamental and applied biological sciences. Psychology Genetic engineering Genetic technics Methods. Procedures. Technologies Plasmids Vectors (cloning, transfer, expression). Insertion sequences and transposons |
| title | A Method for the Re-isolation of an Autonomously Replicating Plasmid from Aspergillus Transformants |
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