Cloning and expression of a beta tubulin gene of Physarum polycephalum
A β tubulin gene of Physarum polycephalum has been isolated from a genomic library in the phage EMBL4. Southern‐blot hybridization to genomic DNA indicates that the cloned DNA is derived from the betB1 locus of the β tubulin gene family. A tubulin‐specific subfragment of the phage DNA was used as a...
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| Veröffentlicht in: | European journal of biochemistry 1988-06, Vol.174 (3), p.491-495 |
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| container_title | European journal of biochemistry |
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| creator | Werenskiold, A.K Poetsch, B Haugli, F |
| description | A β tubulin gene of Physarum polycephalum has been isolated from a genomic library in the phage EMBL4. Southern‐blot hybridization to genomic DNA indicates that the cloned DNA is derived from the betB1 locus of the β tubulin gene family. A tubulin‐specific subfragment of the phage DNA was used as a hybridization probe to construct a restriction map of the betB1 locus. The probe consisted of the almost complete coding region of the 5′ half of the tubulin gene, interrupted by one intron. The derived amino acid sequence of this subclone deviates from the protein sequence for Physarum amoebal β tubulin (amino acids 4–207) in two of 207 amino acids. We used both recA and recBC sbcB bacterial host strains, which have been recommended for cloning of instability‐conferring sequences of the Physarum genome, but were unable to subclone the 3′ part of the gene from the phage DNA. Primer‐extension analysis indicates that the betB gene is expressed in the vegetatively proliferating amoebal and plasmodial stages of the life cycle as well as in differentiating (sporulating) plasmodia. |
| doi_str_mv | 10.1111/j.1432-1033.1988.tb14125.x |
| format | Article |
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Southern‐blot hybridization to genomic DNA indicates that the cloned DNA is derived from the betB1 locus of the β tubulin gene family. A tubulin‐specific subfragment of the phage DNA was used as a hybridization probe to construct a restriction map of the betB1 locus. The probe consisted of the almost complete coding region of the 5′ half of the tubulin gene, interrupted by one intron. The derived amino acid sequence of this subclone deviates from the protein sequence for Physarum amoebal β tubulin (amino acids 4–207) in two of 207 amino acids. We used both recA and recBC sbcB bacterial host strains, which have been recommended for cloning of instability‐conferring sequences of the Physarum genome, but were unable to subclone the 3′ part of the gene from the phage DNA. Primer‐extension analysis indicates that the betB gene is expressed in the vegetatively proliferating amoebal and plasmodial stages of the life cycle as well as in differentiating (sporulating) plasmodia.</description><identifier>ISSN: 0014-2956</identifier><identifier>EISSN: 1432-1033</identifier><identifier>DOI: 10.1111/j.1432-1033.1988.tb14125.x</identifier><identifier>PMID: 3391166</identifier><identifier>CODEN: EJBCAI</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Amino Acid Sequence ; Base Sequence ; Biological and medical sciences ; Biotechnology ; cloning ; Cloning, Molecular ; DNA - analysis ; DNA, Recombinant ; Fundamental and applied biological sciences. Psychology ; gene expression ; Genes ; Genetic engineering ; Genetic technics ; genome ; hybridization ; Methods. Procedures. Technologies ; Molecular cloning ; Molecular Sequence Data ; Nucleic Acid Hybridization ; Physarum - genetics ; Physarum polycephalum ; sporulation ; Tubulin - genetics</subject><ispartof>European journal of biochemistry, 1988-06, Vol.174 (3), p.491-495</ispartof><rights>1988 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4541-e97d47a77199471cd9efdf5623550bb6e7c5004e51188012ea965956ef7087ba3</citedby><cites>FETCH-LOGICAL-c4541-e97d47a77199471cd9efdf5623550bb6e7c5004e51188012ea965956ef7087ba3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7833940$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3391166$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Werenskiold, A.K</creatorcontrib><creatorcontrib>Poetsch, B</creatorcontrib><creatorcontrib>Haugli, F</creatorcontrib><title>Cloning and expression of a beta tubulin gene of Physarum polycephalum</title><title>European journal of biochemistry</title><addtitle>Eur J Biochem</addtitle><description>A β tubulin gene of Physarum polycephalum has been isolated from a genomic library in the phage EMBL4. Southern‐blot hybridization to genomic DNA indicates that the cloned DNA is derived from the betB1 locus of the β tubulin gene family. A tubulin‐specific subfragment of the phage DNA was used as a hybridization probe to construct a restriction map of the betB1 locus. The probe consisted of the almost complete coding region of the 5′ half of the tubulin gene, interrupted by one intron. The derived amino acid sequence of this subclone deviates from the protein sequence for Physarum amoebal β tubulin (amino acids 4–207) in two of 207 amino acids. We used both recA and recBC sbcB bacterial host strains, which have been recommended for cloning of instability‐conferring sequences of the Physarum genome, but were unable to subclone the 3′ part of the gene from the phage DNA. Primer‐extension analysis indicates that the betB gene is expressed in the vegetatively proliferating amoebal and plasmodial stages of the life cycle as well as in differentiating (sporulating) plasmodia.</description><subject>Amino Acid Sequence</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>cloning</subject><subject>Cloning, Molecular</subject><subject>DNA - analysis</subject><subject>DNA, Recombinant</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>gene expression</subject><subject>Genes</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>genome</subject><subject>hybridization</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular cloning</subject><subject>Molecular Sequence Data</subject><subject>Nucleic Acid Hybridization</subject><subject>Physarum - genetics</subject><subject>Physarum polycephalum</subject><subject>sporulation</subject><subject>Tubulin - genetics</subject><issn>0014-2956</issn><issn>1432-1033</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1988</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkktr3DAUhUVpSSdJf0KpKSU7O7rWy-qmtEOmDQRaSLIWsn098SA_Ko3pzL-vzJjZlmoj0PnOfRxEyEegGcRzu8uAszwFylgGuiiyfQkccpEdXpHVWXpNVpQCT3Mt5FtyGcKOUiq1VBfkgjENIOWKbNZu6Nt-m9i-TvAwegyhHfpkaBKblLi3yX4qJ9f2yRZ7nJ9_vRyD9VOXjIM7Vji-WDd11-RNY13Ad8t9RZ43d0_rH-nDz-_3668PacUFhxS1qrmySoHWXEFVa2zqRsicCUHLUqKqBKUcBUBRUMjRaini-NgoWqjSsityc6o7-uH3hGFvujZU6JztcZiCUUWuJVPwTxC4VlxSGcHPJ7DyQwgeGzP6trP-aICaOW2zM3OkZo7UzGmbJW1ziOb3S5ep7LA-W5d4o_5p0W2orGu87as2nDFVRJDTiH05YX9ah8f_GMBs7r49cj2v--FUobGDsVsfmzw_5hRY_ABU8mj9CxQmo9E</recordid><startdate>19880615</startdate><enddate>19880615</enddate><creator>Werenskiold, A.K</creator><creator>Poetsch, B</creator><creator>Haugli, F</creator><general>Blackwell Publishing Ltd</general><general>Blackwell</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>19880615</creationdate><title>Cloning and expression of a beta tubulin gene of Physarum polycephalum</title><author>Werenskiold, A.K ; Poetsch, B ; Haugli, F</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4541-e97d47a77199471cd9efdf5623550bb6e7c5004e51188012ea965956ef7087ba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1988</creationdate><topic>Amino Acid Sequence</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>cloning</topic><topic>Cloning, Molecular</topic><topic>DNA - analysis</topic><topic>DNA, Recombinant</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>gene expression</topic><topic>Genes</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>genome</topic><topic>hybridization</topic><topic>Methods. Procedures. Technologies</topic><topic>Molecular cloning</topic><topic>Molecular Sequence Data</topic><topic>Nucleic Acid Hybridization</topic><topic>Physarum - genetics</topic><topic>Physarum polycephalum</topic><topic>sporulation</topic><topic>Tubulin - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Werenskiold, A.K</creatorcontrib><creatorcontrib>Poetsch, B</creatorcontrib><creatorcontrib>Haugli, F</creatorcontrib><collection>AGRIS</collection><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>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>European journal of biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Werenskiold, A.K</au><au>Poetsch, B</au><au>Haugli, F</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning and expression of a beta tubulin gene of Physarum polycephalum</atitle><jtitle>European journal of biochemistry</jtitle><addtitle>Eur J Biochem</addtitle><date>1988-06-15</date><risdate>1988</risdate><volume>174</volume><issue>3</issue><spage>491</spage><epage>495</epage><pages>491-495</pages><issn>0014-2956</issn><eissn>1432-1033</eissn><coden>EJBCAI</coden><abstract>A β tubulin gene of Physarum polycephalum has been isolated from a genomic library in the phage EMBL4. Southern‐blot hybridization to genomic DNA indicates that the cloned DNA is derived from the betB1 locus of the β tubulin gene family. A tubulin‐specific subfragment of the phage DNA was used as a hybridization probe to construct a restriction map of the betB1 locus. The probe consisted of the almost complete coding region of the 5′ half of the tubulin gene, interrupted by one intron. The derived amino acid sequence of this subclone deviates from the protein sequence for Physarum amoebal β tubulin (amino acids 4–207) in two of 207 amino acids. We used both recA and recBC sbcB bacterial host strains, which have been recommended for cloning of instability‐conferring sequences of the Physarum genome, but were unable to subclone the 3′ part of the gene from the phage DNA. Primer‐extension analysis indicates that the betB gene is expressed in the vegetatively proliferating amoebal and plasmodial stages of the life cycle as well as in differentiating (sporulating) plasmodia.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>3391166</pmid><doi>10.1111/j.1432-1033.1988.tb14125.x</doi><tpages>5</tpages></addata></record> |
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| ispartof | European journal of biochemistry, 1988-06, Vol.174 (3), p.491-495 |
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| subjects | Amino Acid Sequence Base Sequence Biological and medical sciences Biotechnology cloning Cloning, Molecular DNA - analysis DNA, Recombinant Fundamental and applied biological sciences. Psychology gene expression Genes Genetic engineering Genetic technics genome hybridization Methods. Procedures. Technologies Molecular cloning Molecular Sequence Data Nucleic Acid Hybridization Physarum - genetics Physarum polycephalum sporulation Tubulin - genetics |
| title | Cloning and expression of a beta tubulin gene of Physarum polycephalum |
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