Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene
We cloned complementary DNA (cDNA) corresponding to protein_id 10767 using transcripts extracted from a culture expressing ligninolytic enzymes, which is registered as a Ca2+-binding protein in the Phanerochaete chrysosporium whole genome database. We used our results to correct an amino acid sequen...
Gespeichert in:
Veröffentlicht in: | Mycoscience 2012, Vol.54(3), pp.241-246 |
---|---|
Hauptverfasser: | , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
container_end_page | 246 |
---|---|
container_issue | 3 |
container_start_page | 241 |
container_title | Mycoscience |
container_volume | 54 |
creator | Sakamoto, Takaiku Honda, Yoichi Kameshita, Isamu Suzuki, Kazumi Irie, Toshikazu |
description | We cloned complementary DNA (cDNA) corresponding to protein_id 10767 using transcripts extracted from a culture expressing ligninolytic enzymes, which is registered as a Ca2+-binding protein in the Phanerochaete chrysosporium whole genome database. We used our results to correct an amino acid sequence present in the database. Phylogenetic and southern blot analyses of this re-predicted sequence suggest that this is the sole gene encoding calmodulin (CaM) in this fungus. A P. chrysosporium CaM protein (PcCaM) fused to a His-tag was expressed in Escherichia coli. The His-PcCaM showed appropriate CaM features.
▶ We isolated cDNA of calmodulin (CaM) from Phanerochaete chrysosporium. ▶ The sequence corrected the coding region predicted in whole genome database. ▶ Southern blot analysis suggested the gene is the sole CaM gene in the fungus. ▶ The CaM gene was expressed in Escherichia coli. ▶ The recombinant protein showed appropriate CaM features. |
doi_str_mv | 10.1016/j.myc.2012.09.017 |
format | Article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1790939614</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S1340354012000368</els_id><sourcerecordid>1790939614</sourcerecordid><originalsourceid>FETCH-LOGICAL-c486t-415b07d080d80de21a7911331141f1c28e27e0d6a201846328d80925202785703</originalsourceid><addsrcrecordid>eNp9kEtLxDAUhYso-PwB7rp003pvmr5wJYOPAUUXCroKMb0zzdA2Y9IR5997y4hLISSB-53DuSeKzhFSBCwuV2m_NakAFCnUKWC5Fx1hgVUicpnv8z-TkGS5hMPoOIQVMFFk4ih6mwfX6dG6IdZDE7c0knedW7pNiOl77SmEaeYW8dhS_Nzqgeem1czFpvXb4MLaebvpY6O73jWbzg7xkgY6jQ4Wugt09vueRK-3Ny-z--Th6W4-u35IjKyKMZGYf0DZQAUNHxKoyxoxyxAlLtCIikRJ0BSaV6skR66Yq0UuQJRVXkJ2El3sfNfefW4ojKq3wVDXcVJeQmFZQ53VBUpGcYca70LwtFBrb3vttwpBTS2qleIW1dSiglpxR6y53WlWYdRL-lNoP1rT0cS7YKzKpcqm6_F9lksh8Q_grryigY2udkbEZXxZ8oplNBhqrCczqsbZf2L8ABBck4w</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1790939614</pqid></control><display><type>article</type><title>Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene</title><source>J-STAGE Free</source><creator>Sakamoto, Takaiku ; Honda, Yoichi ; Kameshita, Isamu ; Suzuki, Kazumi ; Irie, Toshikazu</creator><creatorcontrib>Sakamoto, Takaiku ; Honda, Yoichi ; Kameshita, Isamu ; Suzuki, Kazumi ; Irie, Toshikazu</creatorcontrib><description>We cloned complementary DNA (cDNA) corresponding to protein_id 10767 using transcripts extracted from a culture expressing ligninolytic enzymes, which is registered as a Ca2+-binding protein in the Phanerochaete chrysosporium whole genome database. We used our results to correct an amino acid sequence present in the database. Phylogenetic and southern blot analyses of this re-predicted sequence suggest that this is the sole gene encoding calmodulin (CaM) in this fungus. A P. chrysosporium CaM protein (PcCaM) fused to a His-tag was expressed in Escherichia coli. The His-PcCaM showed appropriate CaM features.
▶ We isolated cDNA of calmodulin (CaM) from Phanerochaete chrysosporium. ▶ The sequence corrected the coding region predicted in whole genome database. ▶ Southern blot analysis suggested the gene is the sole CaM gene in the fungus. ▶ The CaM gene was expressed in Escherichia coli. ▶ The recombinant protein showed appropriate CaM features.</description><identifier>ISSN: 1340-3540</identifier><identifier>EISSN: 1618-2545</identifier><identifier>DOI: 10.1016/j.myc.2012.09.017</identifier><language>eng</language><publisher>Elsevier B.V</publisher><subject>Basidiomycete ; Electrophoretic migration shift ; Escherichia coli ; Phanerochaete chrysosporium ; Pull-down analysis ; RACE analysis</subject><ispartof>Mycoscience, 2012, Vol.54(3), pp.241-246</ispartof><rights>2012 The Mycological Society of Japan</rights><rights>2012, by The Mycological Society of Japan</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c486t-415b07d080d80de21a7911331141f1c28e27e0d6a201846328d80925202785703</citedby><cites>FETCH-LOGICAL-c486t-415b07d080d80de21a7911331141f1c28e27e0d6a201846328d80925202785703</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,1883,27924,27925</link.rule.ids></links><search><creatorcontrib>Sakamoto, Takaiku</creatorcontrib><creatorcontrib>Honda, Yoichi</creatorcontrib><creatorcontrib>Kameshita, Isamu</creatorcontrib><creatorcontrib>Suzuki, Kazumi</creatorcontrib><creatorcontrib>Irie, Toshikazu</creatorcontrib><title>Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene</title><title>Mycoscience</title><description>We cloned complementary DNA (cDNA) corresponding to protein_id 10767 using transcripts extracted from a culture expressing ligninolytic enzymes, which is registered as a Ca2+-binding protein in the Phanerochaete chrysosporium whole genome database. We used our results to correct an amino acid sequence present in the database. Phylogenetic and southern blot analyses of this re-predicted sequence suggest that this is the sole gene encoding calmodulin (CaM) in this fungus. A P. chrysosporium CaM protein (PcCaM) fused to a His-tag was expressed in Escherichia coli. The His-PcCaM showed appropriate CaM features.
▶ We isolated cDNA of calmodulin (CaM) from Phanerochaete chrysosporium. ▶ The sequence corrected the coding region predicted in whole genome database. ▶ Southern blot analysis suggested the gene is the sole CaM gene in the fungus. ▶ The CaM gene was expressed in Escherichia coli. ▶ The recombinant protein showed appropriate CaM features.</description><subject>Basidiomycete</subject><subject>Electrophoretic migration shift</subject><subject>Escherichia coli</subject><subject>Phanerochaete chrysosporium</subject><subject>Pull-down analysis</subject><subject>RACE analysis</subject><issn>1340-3540</issn><issn>1618-2545</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><recordid>eNp9kEtLxDAUhYso-PwB7rp003pvmr5wJYOPAUUXCroKMb0zzdA2Y9IR5997y4hLISSB-53DuSeKzhFSBCwuV2m_NakAFCnUKWC5Fx1hgVUicpnv8z-TkGS5hMPoOIQVMFFk4ih6mwfX6dG6IdZDE7c0knedW7pNiOl77SmEaeYW8dhS_Nzqgeem1czFpvXb4MLaebvpY6O73jWbzg7xkgY6jQ4Wugt09vueRK-3Ny-z--Th6W4-u35IjKyKMZGYf0DZQAUNHxKoyxoxyxAlLtCIikRJ0BSaV6skR66Yq0UuQJRVXkJ2El3sfNfefW4ojKq3wVDXcVJeQmFZQ53VBUpGcYca70LwtFBrb3vttwpBTS2qleIW1dSiglpxR6y53WlWYdRL-lNoP1rT0cS7YKzKpcqm6_F9lksh8Q_grryigY2udkbEZXxZ8oplNBhqrCczqsbZf2L8ABBck4w</recordid><startdate>20130501</startdate><enddate>20130501</enddate><creator>Sakamoto, Takaiku</creator><creator>Honda, Yoichi</creator><creator>Kameshita, Isamu</creator><creator>Suzuki, Kazumi</creator><creator>Irie, Toshikazu</creator><general>Elsevier B.V</general><general>The Mycological Society of Japan</general><scope>AAYXX</scope><scope>CITATION</scope><scope>M7N</scope></search><sort><creationdate>20130501</creationdate><title>Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene</title><author>Sakamoto, Takaiku ; Honda, Yoichi ; Kameshita, Isamu ; Suzuki, Kazumi ; Irie, Toshikazu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c486t-415b07d080d80de21a7911331141f1c28e27e0d6a201846328d80925202785703</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Basidiomycete</topic><topic>Electrophoretic migration shift</topic><topic>Escherichia coli</topic><topic>Phanerochaete chrysosporium</topic><topic>Pull-down analysis</topic><topic>RACE analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sakamoto, Takaiku</creatorcontrib><creatorcontrib>Honda, Yoichi</creatorcontrib><creatorcontrib>Kameshita, Isamu</creatorcontrib><creatorcontrib>Suzuki, Kazumi</creatorcontrib><creatorcontrib>Irie, Toshikazu</creatorcontrib><collection>CrossRef</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><jtitle>Mycoscience</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sakamoto, Takaiku</au><au>Honda, Yoichi</au><au>Kameshita, Isamu</au><au>Suzuki, Kazumi</au><au>Irie, Toshikazu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene</atitle><jtitle>Mycoscience</jtitle><date>2013-05-01</date><risdate>2013</risdate><volume>54</volume><issue>3</issue><spage>241</spage><epage>246</epage><pages>241-246</pages><issn>1340-3540</issn><eissn>1618-2545</eissn><abstract>We cloned complementary DNA (cDNA) corresponding to protein_id 10767 using transcripts extracted from a culture expressing ligninolytic enzymes, which is registered as a Ca2+-binding protein in the Phanerochaete chrysosporium whole genome database. We used our results to correct an amino acid sequence present in the database. Phylogenetic and southern blot analyses of this re-predicted sequence suggest that this is the sole gene encoding calmodulin (CaM) in this fungus. A P. chrysosporium CaM protein (PcCaM) fused to a His-tag was expressed in Escherichia coli. The His-PcCaM showed appropriate CaM features.
▶ We isolated cDNA of calmodulin (CaM) from Phanerochaete chrysosporium. ▶ The sequence corrected the coding region predicted in whole genome database. ▶ Southern blot analysis suggested the gene is the sole CaM gene in the fungus. ▶ The CaM gene was expressed in Escherichia coli. ▶ The recombinant protein showed appropriate CaM features.</abstract><pub>Elsevier B.V</pub><doi>10.1016/j.myc.2012.09.017</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 1340-3540 |
ispartof | Mycoscience, 2012, Vol.54(3), pp.241-246 |
issn | 1340-3540 1618-2545 |
language | eng |
recordid | cdi_proquest_miscellaneous_1790939614 |
source | J-STAGE Free |
subjects | Basidiomycete Electrophoretic migration shift Escherichia coli Phanerochaete chrysosporium Pull-down analysis RACE analysis |
title | Isolation and heterologous expression of the Phanerochaete chrysosporium calmodulin gene |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-04T12%3A03%3A14IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Isolation%20and%20heterologous%20expression%20of%20the%20Phanerochaete%20chrysosporium%20calmodulin%20gene&rft.jtitle=Mycoscience&rft.au=Sakamoto,%20Takaiku&rft.date=2013-05-01&rft.volume=54&rft.issue=3&rft.spage=241&rft.epage=246&rft.pages=241-246&rft.issn=1340-3540&rft.eissn=1618-2545&rft_id=info:doi/10.1016/j.myc.2012.09.017&rft_dat=%3Cproquest_cross%3E1790939614%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1790939614&rft_id=info:pmid/&rft_els_id=S1340354012000368&rfr_iscdi=true |