A recyclable Candida albicans URA3 cassette for PCR product‐directed gene disruptions
For some time, gene disruptions in Candida albicans have been made with the hisG‐URA3‐hisG (‘Ura‐blaster’) cassette, which can be re‐used in successive transformations of a single strain after homologous excision of URA3. However, the hisG repeats are too large for efficient PCR amplification of the...
Gespeichert in:
| Veröffentlicht in: | Yeast (Chichester, England) England), 2000-01, Vol.16 (1), p.65-70 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 70 |
|---|---|
| container_issue | 1 |
| container_start_page | 65 |
| container_title | Yeast (Chichester, England) |
| container_volume | 16 |
| creator | Bryce Wilson, R. Davis, Dana Enloe, Brian M. Mitchell, Aaron P. |
| description | For some time, gene disruptions in Candida albicans have been made with the hisG‐URA3‐hisG (‘Ura‐blaster’) cassette, which can be re‐used in successive transformations of a single strain after homologous excision of URA3. However, the hisG repeats are too large for efficient PCR amplification of the entire cassette, so it cannot be used for PCR product‐directed gene disruptions. We describe here a gene disruption cassette, URA3‐dpl200, with 200 bp flanking repeats that permit efficient PCR amplification. After transformation and integration to produce both arg5::URA3‐dpl200 and rim101::URA3‐dpl200 alleles, we find that arg5::dpl200 and rim101::dpl200 segregants, respectively, can be obtained. We have used the cassette to create rim101::dpl200/rim101::URA3‐dpl200 mutants exclusively through PCR product‐directed disruption. Copyright © 2000 John Wiley & Sons, Ltd. |
| doi_str_mv | 10.1002/(SICI)1097-0061(20000115)16:1<65::AID-YEA508>3.0.CO;2-M |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_70817970</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>70817970</sourcerecordid><originalsourceid>FETCH-LOGICAL-p2898-d71a0f322392b2b5c81bffa2c11c48abdb700010e5f912256ea37bcaddaf11183</originalsourceid><addsrcrecordid>eNpNkc9u1EAMxkeIim4Lr4DmhNpDFnumySQLoopCKSu1WlSooCdr_gUFZbPbTKJqbzxCn7FP0qS7Rfhiyf5s2d-PsVOEKQKI90ff58X8GCFTEUCCRwKGQIyPMZnhxySezfL55-jmLI8h_SSnMC0WH0R0-YJN_s28ZBNQJ1kUg_y1zw5C-PO0QaSv2D5CIkCpZMJ-5rz1dmNrbWrPC924ymmua1NZ3QR-fZVLbnUIvus8L1ct_1Zc8XW7cr3tHv7eu2qY7rzjv33juatC26-7atWE12yv1HXwb3b5kF1_OftRfI0uFufzIr-I1iLN0sgp1FBKIWQmjDCxTdGUpRYW0Z6k2jijxsfBx2WGQsSJ11IZq53TJSKm8pC92-4dbrrtfehoWQXr61o3ftUHUpCiyhQMwrc7YW-W3tG6rZa63dCzFYPgZiu4q2q_-a9PIxEagdBoLo3m0jMQwoSQkpgGHrTlQZKAigUJutxV5COEJIPA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>70817970</pqid></control><display><type>article</type><title>A recyclable Candida albicans URA3 cassette for PCR product‐directed gene disruptions</title><source>MEDLINE</source><source>Wiley Online Library</source><source>Wiley Online Library Journals</source><source>PubMed Central</source><source>EZB Electronic Journals Library</source><source>PubMed Central Open Access</source><creator>Bryce Wilson, R. ; Davis, Dana ; Enloe, Brian M. ; Mitchell, Aaron P.</creator><creatorcontrib>Bryce Wilson, R. ; Davis, Dana ; Enloe, Brian M. ; Mitchell, Aaron P.</creatorcontrib><description>For some time, gene disruptions in Candida albicans have been made with the hisG‐URA3‐hisG (‘Ura‐blaster’) cassette, which can be re‐used in successive transformations of a single strain after homologous excision of URA3. However, the hisG repeats are too large for efficient PCR amplification of the entire cassette, so it cannot be used for PCR product‐directed gene disruptions. We describe here a gene disruption cassette, URA3‐dpl200, with 200 bp flanking repeats that permit efficient PCR amplification. After transformation and integration to produce both arg5::URA3‐dpl200 and rim101::URA3‐dpl200 alleles, we find that arg5::dpl200 and rim101::dpl200 segregants, respectively, can be obtained. We have used the cassette to create rim101::dpl200/rim101::URA3‐dpl200 mutants exclusively through PCR product‐directed disruption. Copyright © 2000 John Wiley & Sons, Ltd.</description><identifier>ISSN: 0749-503X</identifier><identifier>EISSN: 1097-0061</identifier><identifier>DOI: 10.1002/(SICI)1097-0061(20000115)16:1<65::AID-YEA508>3.0.CO;2-M</identifier><identifier>PMID: 10620776</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Candida albicans ; Candida albicans - genetics ; Fungal Proteins - genetics ; gene disruption ; Polymerase Chain Reaction ; Transformation, Genetic ; Ura‐blaster</subject><ispartof>Yeast (Chichester, England), 2000-01, Vol.16 (1), p.65-70</ispartof><rights>Copyright © 2000 John Wiley & Sons, Ltd.</rights><rights>Copyright 2000 John Wiley & Sons, Ltd.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2F%28SICI%291097-0061%2820000115%2916%3A1%3C65%3A%3AAID-YEA508%3E3.0.CO%3B2-M$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2F%28SICI%291097-0061%2820000115%2916%3A1%3C65%3A%3AAID-YEA508%3E3.0.CO%3B2-M$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,1427,27901,27902,45550,45551,46384,46808</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10620776$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bryce Wilson, R.</creatorcontrib><creatorcontrib>Davis, Dana</creatorcontrib><creatorcontrib>Enloe, Brian M.</creatorcontrib><creatorcontrib>Mitchell, Aaron P.</creatorcontrib><title>A recyclable Candida albicans URA3 cassette for PCR product‐directed gene disruptions</title><title>Yeast (Chichester, England)</title><addtitle>Yeast</addtitle><description>For some time, gene disruptions in Candida albicans have been made with the hisG‐URA3‐hisG (‘Ura‐blaster’) cassette, which can be re‐used in successive transformations of a single strain after homologous excision of URA3. However, the hisG repeats are too large for efficient PCR amplification of the entire cassette, so it cannot be used for PCR product‐directed gene disruptions. We describe here a gene disruption cassette, URA3‐dpl200, with 200 bp flanking repeats that permit efficient PCR amplification. After transformation and integration to produce both arg5::URA3‐dpl200 and rim101::URA3‐dpl200 alleles, we find that arg5::dpl200 and rim101::dpl200 segregants, respectively, can be obtained. We have used the cassette to create rim101::dpl200/rim101::URA3‐dpl200 mutants exclusively through PCR product‐directed disruption. Copyright © 2000 John Wiley & Sons, Ltd.</description><subject>Candida albicans</subject><subject>Candida albicans - genetics</subject><subject>Fungal Proteins - genetics</subject><subject>gene disruption</subject><subject>Polymerase Chain Reaction</subject><subject>Transformation, Genetic</subject><subject>Ura‐blaster</subject><issn>0749-503X</issn><issn>1097-0061</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpNkc9u1EAMxkeIim4Lr4DmhNpDFnumySQLoopCKSu1WlSooCdr_gUFZbPbTKJqbzxCn7FP0qS7Rfhiyf5s2d-PsVOEKQKI90ff58X8GCFTEUCCRwKGQIyPMZnhxySezfL55-jmLI8h_SSnMC0WH0R0-YJN_s28ZBNQJ1kUg_y1zw5C-PO0QaSv2D5CIkCpZMJ-5rz1dmNrbWrPC924ymmua1NZ3QR-fZVLbnUIvus8L1ct_1Zc8XW7cr3tHv7eu2qY7rzjv33juatC26-7atWE12yv1HXwb3b5kF1_OftRfI0uFufzIr-I1iLN0sgp1FBKIWQmjDCxTdGUpRYW0Z6k2jijxsfBx2WGQsSJ11IZq53TJSKm8pC92-4dbrrtfehoWQXr61o3ftUHUpCiyhQMwrc7YW-W3tG6rZa63dCzFYPgZiu4q2q_-a9PIxEagdBoLo3m0jMQwoSQkpgGHrTlQZKAigUJutxV5COEJIPA</recordid><startdate>20000115</startdate><enddate>20000115</enddate><creator>Bryce Wilson, R.</creator><creator>Davis, Dana</creator><creator>Enloe, Brian M.</creator><creator>Mitchell, Aaron P.</creator><general>John Wiley & Sons, Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>20000115</creationdate><title>A recyclable Candida albicans URA3 cassette for PCR product‐directed gene disruptions</title><author>Bryce Wilson, R. ; Davis, Dana ; Enloe, Brian M. ; Mitchell, Aaron P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p2898-d71a0f322392b2b5c81bffa2c11c48abdb700010e5f912256ea37bcaddaf11183</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Candida albicans</topic><topic>Candida albicans - genetics</topic><topic>Fungal Proteins - genetics</topic><topic>gene disruption</topic><topic>Polymerase Chain Reaction</topic><topic>Transformation, Genetic</topic><topic>Ura‐blaster</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bryce Wilson, R.</creatorcontrib><creatorcontrib>Davis, Dana</creatorcontrib><creatorcontrib>Enloe, Brian M.</creatorcontrib><creatorcontrib>Mitchell, Aaron P.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Yeast (Chichester, England)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bryce Wilson, R.</au><au>Davis, Dana</au><au>Enloe, Brian M.</au><au>Mitchell, Aaron P.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A recyclable Candida albicans URA3 cassette for PCR product‐directed gene disruptions</atitle><jtitle>Yeast (Chichester, England)</jtitle><addtitle>Yeast</addtitle><date>2000-01-15</date><risdate>2000</risdate><volume>16</volume><issue>1</issue><spage>65</spage><epage>70</epage><pages>65-70</pages><issn>0749-503X</issn><eissn>1097-0061</eissn><abstract>For some time, gene disruptions in Candida albicans have been made with the hisG‐URA3‐hisG (‘Ura‐blaster’) cassette, which can be re‐used in successive transformations of a single strain after homologous excision of URA3. However, the hisG repeats are too large for efficient PCR amplification of the entire cassette, so it cannot be used for PCR product‐directed gene disruptions. We describe here a gene disruption cassette, URA3‐dpl200, with 200 bp flanking repeats that permit efficient PCR amplification. After transformation and integration to produce both arg5::URA3‐dpl200 and rim101::URA3‐dpl200 alleles, we find that arg5::dpl200 and rim101::dpl200 segregants, respectively, can be obtained. We have used the cassette to create rim101::dpl200/rim101::URA3‐dpl200 mutants exclusively through PCR product‐directed disruption. Copyright © 2000 John Wiley & Sons, Ltd.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>10620776</pmid><doi>10.1002/(SICI)1097-0061(20000115)16:1<65::AID-YEA508>3.0.CO;2-M</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0749-503X |
| ispartof | Yeast (Chichester, England), 2000-01, Vol.16 (1), p.65-70 |
| issn | 0749-503X 1097-0061 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_70817970 |
| source | MEDLINE; Wiley Online Library; Wiley Online Library Journals; PubMed Central; EZB Electronic Journals Library; PubMed Central Open Access |
| subjects | Candida albicans Candida albicans - genetics Fungal Proteins - genetics gene disruption Polymerase Chain Reaction Transformation, Genetic Ura‐blaster |
| title | A recyclable Candida albicans URA3 cassette for PCR product‐directed gene disruptions |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-07T16%3A23%3A59IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20recyclable%20Candida%20albicans%20URA3%20cassette%20for%20PCR%20product%E2%80%90directed%20gene%20disruptions&rft.jtitle=Yeast%20(Chichester,%20England)&rft.au=Bryce%20Wilson,%20R.&rft.date=2000-01-15&rft.volume=16&rft.issue=1&rft.spage=65&rft.epage=70&rft.pages=65-70&rft.issn=0749-503X&rft.eissn=1097-0061&rft_id=info:doi/10.1002/(SICI)1097-0061(20000115)16:1%3C65::AID-YEA508%3E3.0.CO;2-M&rft_dat=%3Cproquest_pubme%3E70817970%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=70817970&rft_id=info:pmid/10620776&rfr_iscdi=true |