Synthesis of the measles virus nucleoprotein in yeast Pichia pastoris and Saccharomyces cerevisiae
The development of a simple, efficient and cost-effective system for generation of measles virus nucleoprotein might help to upgrade reagents for measles serology. The gene encoding measles nucleoprotein was successfully expressed in two different yeast genera, Pichia pastoris and Saccharomyces cere...
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| Veröffentlicht in: | Journal of biotechnology 2004-01, Vol.107 (2), p.115-124 |
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| container_title | Journal of biotechnology |
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| creator | Slibinskas, Rimantas Samuel, Dhanraj Gedvilaite, Alma Staniulis, Juozas Sasnauskas, Kestutis |
| description | The development of a simple, efficient and cost-effective system for generation of measles virus nucleoprotein might help to upgrade reagents for measles serology. The gene encoding measles nucleoprotein was successfully expressed in two different yeast genera,
Pichia pastoris and
Saccharomyces cerevisiae, respectively. Both yeast genera synthesized a high level of nucleoprotein, up to 29 and 18% of total cell protein, in
P. pastoris and
S. cerevisiae, respectively. This protein is one of most abundantly expressed in yeast. After purification nucleocapsid-like particles (NLPs) derived from both yeast genera appeared to be similar to those detected in mammalian cells infected with measles virus. A spontaneous assembly of nucleoprotein into nucleocapsid-like particles in the absence of the viral leader RNA or viral proteins has been shown. Compartmentalisation of recombinant protein into large compact inclusions in the cytoplasm of yeast
S. cerevisiae by green fluorescence protein (GFP) fusion has been demonstrated. Sera from measles patients reacted with the recombinant protein expressed in both yeast genera and a simple diagnostic assay to detect measles IgM could be designed on this basis. |
| doi_str_mv | 10.1016/j.jbiotec.2003.10.018 |
| format | Article |
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Pichia pastoris and
Saccharomyces cerevisiae, respectively. Both yeast genera synthesized a high level of nucleoprotein, up to 29 and 18% of total cell protein, in
P. pastoris and
S. cerevisiae, respectively. This protein is one of most abundantly expressed in yeast. After purification nucleocapsid-like particles (NLPs) derived from both yeast genera appeared to be similar to those detected in mammalian cells infected with measles virus. A spontaneous assembly of nucleoprotein into nucleocapsid-like particles in the absence of the viral leader RNA or viral proteins has been shown. Compartmentalisation of recombinant protein into large compact inclusions in the cytoplasm of yeast
S. cerevisiae by green fluorescence protein (GFP) fusion has been demonstrated. Sera from measles patients reacted with the recombinant protein expressed in both yeast genera and a simple diagnostic assay to detect measles IgM could be designed on this basis.</description><identifier>ISSN: 0168-1656</identifier><identifier>EISSN: 1873-4863</identifier><identifier>DOI: 10.1016/j.jbiotec.2003.10.018</identifier><identifier>PMID: 14711495</identifier><identifier>CODEN: JBITD4</identifier><language>eng</language><publisher>Lausanne: Elsevier B.V</publisher><subject>Biological and medical sciences ; Biotechnology ; Blotting, Western ; Cell Compartmentation ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Expression ; Fundamental and applied biological sciences. Psychology ; Gene Dosage ; Green Fluorescent Proteins ; Humans ; Immunoglobulin M - blood ; Kinetics ; Luminescent Proteins ; Measles ; Measles - virology ; Measles virus ; Measles virus - chemistry ; Measles virus - immunology ; Measles virus - metabolism ; Nucleocapsid-like particles ; Nucleoproteins - biosynthesis ; Nucleoproteins - genetics ; Nucleoproteins - immunology ; Nucleoproteins - isolation & purification ; Nucleoproteins - metabolism ; Nucleoproteins - ultrastructure ; Pichia - metabolism ; Pichia pastoris ; Recombinant Fusion Proteins - biosynthesis ; Recombinant Fusion Proteins - metabolism ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - metabolism ; Spectrometry, Fluorescence</subject><ispartof>Journal of biotechnology, 2004-01, Vol.107 (2), p.115-124</ispartof><rights>2003 Elsevier B.V.</rights><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c422t-5dcc892d42a9c5577eb4b50f50abcf4de90a98d0c9a9ce452df6776c08e875353</citedby><cites>FETCH-LOGICAL-c422t-5dcc892d42a9c5577eb4b50f50abcf4de90a98d0c9a9ce452df6776c08e875353</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jbiotec.2003.10.018$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15383682$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14711495$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Slibinskas, Rimantas</creatorcontrib><creatorcontrib>Samuel, Dhanraj</creatorcontrib><creatorcontrib>Gedvilaite, Alma</creatorcontrib><creatorcontrib>Staniulis, Juozas</creatorcontrib><creatorcontrib>Sasnauskas, Kestutis</creatorcontrib><title>Synthesis of the measles virus nucleoprotein in yeast Pichia pastoris and Saccharomyces cerevisiae</title><title>Journal of biotechnology</title><addtitle>J Biotechnol</addtitle><description>The development of a simple, efficient and cost-effective system for generation of measles virus nucleoprotein might help to upgrade reagents for measles serology. The gene encoding measles nucleoprotein was successfully expressed in two different yeast genera,
Pichia pastoris and
Saccharomyces cerevisiae, respectively. Both yeast genera synthesized a high level of nucleoprotein, up to 29 and 18% of total cell protein, in
P. pastoris and
S. cerevisiae, respectively. This protein is one of most abundantly expressed in yeast. After purification nucleocapsid-like particles (NLPs) derived from both yeast genera appeared to be similar to those detected in mammalian cells infected with measles virus. A spontaneous assembly of nucleoprotein into nucleocapsid-like particles in the absence of the viral leader RNA or viral proteins has been shown. Compartmentalisation of recombinant protein into large compact inclusions in the cytoplasm of yeast
S. cerevisiae by green fluorescence protein (GFP) fusion has been demonstrated. Sera from measles patients reacted with the recombinant protein expressed in both yeast genera and a simple diagnostic assay to detect measles IgM could be designed on this basis.</description><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Blotting, Western</subject><subject>Cell Compartmentation</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Escherichia coli</subject><subject>Expression</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Dosage</subject><subject>Green Fluorescent Proteins</subject><subject>Humans</subject><subject>Immunoglobulin M - blood</subject><subject>Kinetics</subject><subject>Luminescent Proteins</subject><subject>Measles</subject><subject>Measles - virology</subject><subject>Measles virus</subject><subject>Measles virus - chemistry</subject><subject>Measles virus - immunology</subject><subject>Measles virus - metabolism</subject><subject>Nucleocapsid-like particles</subject><subject>Nucleoproteins - biosynthesis</subject><subject>Nucleoproteins - genetics</subject><subject>Nucleoproteins - immunology</subject><subject>Nucleoproteins - isolation & purification</subject><subject>Nucleoproteins - metabolism</subject><subject>Nucleoproteins - ultrastructure</subject><subject>Pichia - metabolism</subject><subject>Pichia pastoris</subject><subject>Recombinant Fusion Proteins - biosynthesis</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - metabolism</subject><subject>Spectrometry, Fluorescence</subject><issn>0168-1656</issn><issn>1873-4863</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1r3DAQhkVpSbZpfkKLLu3N25Et2fKplJB-QKCFpGchj8esFn9sJXth_31mWUOOAYGGmeedGeYV4qOCrQJVft1v902YZsJtDlBwbgvKvhEbZasi07Ys3ooNczZTpSmvxfuU9gCga6OuxLXSlVIcb0TzeBrnHaWQ5NRJjuRAPvWU5DHEJclxwZ6mQ-RJYZT8Tlye5d-Au-DlgeMpstaPrXz0iDsfp-GELEeKdAwpePog3nW-T3S7_jfi34_7p7tf2cOfn7_vvj9kqPN8zkyLaOu81bmv0ZiqokY3BjoDvsFOt1SDr20LWHOdtMnbrqyqEsGSrUxhihvx5dKXt_2_UJrdEBJS3_uRpiU5C2A1gHoVVHWutFE1g-YCYpxSitS5QwyDjyenwJ1dcHu3uuDOLpzT7ALrPq0Dlmag9kW1np2BzyvgE_q-i37EkF44U9iitDlz3y4c8d2OgaJLGGhEakMknF07hVdWeQa2JqnU</recordid><startdate>20040122</startdate><enddate>20040122</enddate><creator>Slibinskas, Rimantas</creator><creator>Samuel, Dhanraj</creator><creator>Gedvilaite, Alma</creator><creator>Staniulis, Juozas</creator><creator>Sasnauskas, Kestutis</creator><general>Elsevier B.V</general><general>Elsevier</general><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>7QO</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20040122</creationdate><title>Synthesis of the measles virus nucleoprotein in yeast Pichia pastoris and Saccharomyces cerevisiae</title><author>Slibinskas, Rimantas ; Samuel, Dhanraj ; Gedvilaite, Alma ; Staniulis, Juozas ; Sasnauskas, Kestutis</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c422t-5dcc892d42a9c5577eb4b50f50abcf4de90a98d0c9a9ce452df6776c08e875353</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Blotting, Western</topic><topic>Cell Compartmentation</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Escherichia coli</topic><topic>Expression</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Dosage</topic><topic>Green Fluorescent Proteins</topic><topic>Humans</topic><topic>Immunoglobulin M - blood</topic><topic>Kinetics</topic><topic>Luminescent Proteins</topic><topic>Measles</topic><topic>Measles - virology</topic><topic>Measles virus</topic><topic>Measles virus - chemistry</topic><topic>Measles virus - immunology</topic><topic>Measles virus - metabolism</topic><topic>Nucleocapsid-like particles</topic><topic>Nucleoproteins - biosynthesis</topic><topic>Nucleoproteins - genetics</topic><topic>Nucleoproteins - immunology</topic><topic>Nucleoproteins - isolation & purification</topic><topic>Nucleoproteins - metabolism</topic><topic>Nucleoproteins - ultrastructure</topic><topic>Pichia - metabolism</topic><topic>Pichia pastoris</topic><topic>Recombinant Fusion Proteins - biosynthesis</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Saccharomyces cerevisiae</topic><topic>Saccharomyces cerevisiae - metabolism</topic><topic>Spectrometry, Fluorescence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Slibinskas, Rimantas</creatorcontrib><creatorcontrib>Samuel, Dhanraj</creatorcontrib><creatorcontrib>Gedvilaite, Alma</creatorcontrib><creatorcontrib>Staniulis, Juozas</creatorcontrib><creatorcontrib>Sasnauskas, Kestutis</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>Biotechnology Research Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Slibinskas, Rimantas</au><au>Samuel, Dhanraj</au><au>Gedvilaite, Alma</au><au>Staniulis, Juozas</au><au>Sasnauskas, Kestutis</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Synthesis of the measles virus nucleoprotein in yeast Pichia pastoris and Saccharomyces cerevisiae</atitle><jtitle>Journal of biotechnology</jtitle><addtitle>J Biotechnol</addtitle><date>2004-01-22</date><risdate>2004</risdate><volume>107</volume><issue>2</issue><spage>115</spage><epage>124</epage><pages>115-124</pages><issn>0168-1656</issn><eissn>1873-4863</eissn><coden>JBITD4</coden><abstract>The development of a simple, efficient and cost-effective system for generation of measles virus nucleoprotein might help to upgrade reagents for measles serology. The gene encoding measles nucleoprotein was successfully expressed in two different yeast genera,
Pichia pastoris and
Saccharomyces cerevisiae, respectively. Both yeast genera synthesized a high level of nucleoprotein, up to 29 and 18% of total cell protein, in
P. pastoris and
S. cerevisiae, respectively. This protein is one of most abundantly expressed in yeast. After purification nucleocapsid-like particles (NLPs) derived from both yeast genera appeared to be similar to those detected in mammalian cells infected with measles virus. A spontaneous assembly of nucleoprotein into nucleocapsid-like particles in the absence of the viral leader RNA or viral proteins has been shown. Compartmentalisation of recombinant protein into large compact inclusions in the cytoplasm of yeast
S. cerevisiae by green fluorescence protein (GFP) fusion has been demonstrated. Sera from measles patients reacted with the recombinant protein expressed in both yeast genera and a simple diagnostic assay to detect measles IgM could be designed on this basis.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>14711495</pmid><doi>10.1016/j.jbiotec.2003.10.018</doi><tpages>10</tpages></addata></record> |
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| subjects | Biological and medical sciences Biotechnology Blotting, Western Cell Compartmentation Electrophoresis, Polyacrylamide Gel Escherichia coli Expression Fundamental and applied biological sciences. Psychology Gene Dosage Green Fluorescent Proteins Humans Immunoglobulin M - blood Kinetics Luminescent Proteins Measles Measles - virology Measles virus Measles virus - chemistry Measles virus - immunology Measles virus - metabolism Nucleocapsid-like particles Nucleoproteins - biosynthesis Nucleoproteins - genetics Nucleoproteins - immunology Nucleoproteins - isolation & purification Nucleoproteins - metabolism Nucleoproteins - ultrastructure Pichia - metabolism Pichia pastoris Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae - metabolism Spectrometry, Fluorescence |
| title | Synthesis of the measles virus nucleoprotein in yeast Pichia pastoris and Saccharomyces cerevisiae |
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