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Efficient Transient Expression of Recombinant Proteins in Plants by the Novel pEff Vector Based on the Genome of Potato Virus X
Agroinfiltration of plant leaves with binary vectors carrying a gene of interest within a plant viral vector is a rapid and efficient method for protein production in plants. Previously, we constructed a self-replicating vector, pA7248AMV, based on the genetic elements of potato virus X (PVX), and h...
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| Published in: | Frontiers in plant science 2017-02, Vol.8, p.247-247 |
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| container_title | Frontiers in plant science |
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| creator | Mardanova, Eugenia S Blokhina, Elena A Tsybalova, Liudmila M Peyret, Hadrien Lomonossoff, George P Ravin, Nikolai V |
| description | Agroinfiltration of plant leaves with binary vectors carrying a gene of interest within a plant viral vector is a rapid and efficient method for protein production in plants. Previously, we constructed a self-replicating vector, pA7248AMV, based on the genetic elements of potato virus X (PVX), and have shown that this vector can be used for the expression of recombinant proteins in
. However, this vector is almost 18 kb long and therefore not convenient for genetic manipulation. Furthermore, for efficient expression of the target protein it should be co-agroinfiltrated with an additional binary vector expressing a suppressor of post-transcriptional gene silencing. Here, we improved this expression system by creating the novel pEff vector. Its backbone is about 5 kb shorter than the original vector and it contains an expression cassette for the silencing suppressor, P24, from grapevine leafroll-associated virus-2 alongside PVX genetic elements, thus eliminating the need of co-agroinfiltration. The pEff vector provides green fluorescent protein expression levels of up to 30% of total soluble protein. The novel vector was used for expression of the influenza vaccine candidate, M2eHBc, consisting of an extracellular domain of influenza virus M2 protein (M2e) fused to hepatitis B core antigen. Using the pEff system, M2eHBc was expressed to 5-10% of total soluble protein, several times higher than with original pA7248AMV vector. Plant-produced M2eHBc formed virus-like particles
, as required for its use as a vaccine. The new self-replicating pEff vector could be used for fast and efficient production of various recombinant proteins in plants. |
| doi_str_mv | 10.3389/fpls.2017.00247 |
| format | article |
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. However, this vector is almost 18 kb long and therefore not convenient for genetic manipulation. Furthermore, for efficient expression of the target protein it should be co-agroinfiltrated with an additional binary vector expressing a suppressor of post-transcriptional gene silencing. Here, we improved this expression system by creating the novel pEff vector. Its backbone is about 5 kb shorter than the original vector and it contains an expression cassette for the silencing suppressor, P24, from grapevine leafroll-associated virus-2 alongside PVX genetic elements, thus eliminating the need of co-agroinfiltration. The pEff vector provides green fluorescent protein expression levels of up to 30% of total soluble protein. The novel vector was used for expression of the influenza vaccine candidate, M2eHBc, consisting of an extracellular domain of influenza virus M2 protein (M2e) fused to hepatitis B core antigen. Using the pEff system, M2eHBc was expressed to 5-10% of total soluble protein, several times higher than with original pA7248AMV vector. Plant-produced M2eHBc formed virus-like particles
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. However, this vector is almost 18 kb long and therefore not convenient for genetic manipulation. Furthermore, for efficient expression of the target protein it should be co-agroinfiltrated with an additional binary vector expressing a suppressor of post-transcriptional gene silencing. Here, we improved this expression system by creating the novel pEff vector. Its backbone is about 5 kb shorter than the original vector and it contains an expression cassette for the silencing suppressor, P24, from grapevine leafroll-associated virus-2 alongside PVX genetic elements, thus eliminating the need of co-agroinfiltration. The pEff vector provides green fluorescent protein expression levels of up to 30% of total soluble protein. The novel vector was used for expression of the influenza vaccine candidate, M2eHBc, consisting of an extracellular domain of influenza virus M2 protein (M2e) fused to hepatitis B core antigen. Using the pEff system, M2eHBc was expressed to 5-10% of total soluble protein, several times higher than with original pA7248AMV vector. Plant-produced M2eHBc formed virus-like particles
, as required for its use as a vaccine. The new self-replicating pEff vector could be used for fast and efficient production of various recombinant proteins in plants.</description><subject>Plant Science</subject><issn>1664-462X</issn><issn>1664-462X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><recordid>eNpVkUtPxCAUhYnRqFHX7gxLNzPyagsbEzXjIzE6MWrcEdpeFNOWCp2JrvzrUl9RNtxwDt-9cBDapWTKuVQHtm_ilBFaTAlholhBmzTPxUTk7GH1T72BdmJ8JmllhChVrKMNJpniTIhN9D6z1lUOugHfBtPFz2r22geI0fkOe4tvoPJt6TqTlHnwA7guYtfheZNOIi7f8PAE-MovocF9wuF7qAYf8LGJUOPEGOUz6HwLI27uBzN4fO_CIuKHbbRmTRNh53vfQnens9uT88nl9dnFydHlpBKZGiaZoIJQMLnlZW25tDUDXuaVULxUplBAjQAjVC1AMmZMBoLUgnDLuJRlafgWOvzi9ouyhbpKzwym0X1wrQlv2hun_yude9KPfqkzzqQSRQLsfwOCf1lAHHTrYgVN-gTwi6ipLAqZpSFpsh58WavgYwxgf9tQosfk9JicHpPTn8mlG3t_p_v1_-TEPwAeNpec</recordid><startdate>20170228</startdate><enddate>20170228</enddate><creator>Mardanova, Eugenia S</creator><creator>Blokhina, Elena A</creator><creator>Tsybalova, Liudmila M</creator><creator>Peyret, Hadrien</creator><creator>Lomonossoff, George P</creator><creator>Ravin, Nikolai V</creator><general>Frontiers Media S.A</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20170228</creationdate><title>Efficient Transient Expression of Recombinant Proteins in Plants by the Novel pEff Vector Based on the Genome of Potato Virus X</title><author>Mardanova, Eugenia S ; Blokhina, Elena A ; Tsybalova, Liudmila M ; Peyret, Hadrien ; Lomonossoff, George P ; Ravin, Nikolai V</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c459t-541401ea6f3bdf38fd2e3b6c493b9a79e1a4ea49d4e822aa5e40d403f2388bba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Plant Science</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mardanova, Eugenia S</creatorcontrib><creatorcontrib>Blokhina, Elena A</creatorcontrib><creatorcontrib>Tsybalova, Liudmila M</creatorcontrib><creatorcontrib>Peyret, Hadrien</creatorcontrib><creatorcontrib>Lomonossoff, George P</creatorcontrib><creatorcontrib>Ravin, Nikolai V</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Frontiers in plant science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mardanova, Eugenia S</au><au>Blokhina, Elena A</au><au>Tsybalova, Liudmila M</au><au>Peyret, Hadrien</au><au>Lomonossoff, George P</au><au>Ravin, Nikolai V</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficient Transient Expression of Recombinant Proteins in Plants by the Novel pEff Vector Based on the Genome of Potato Virus X</atitle><jtitle>Frontiers in plant science</jtitle><addtitle>Front Plant Sci</addtitle><date>2017-02-28</date><risdate>2017</risdate><volume>8</volume><spage>247</spage><epage>247</epage><pages>247-247</pages><issn>1664-462X</issn><eissn>1664-462X</eissn><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><notes>This article was submitted to Plant Biotechnology, a section of the journal Frontiers in Plant Science</notes><notes>Edited by: Joachim Hermann Schiemann, Julius Kühn-Institut, Germany</notes><notes>Reviewed by: Sergey Morozov, Moscow State University, Russia; Chiara Lico, Italian National Agency on New Technologies, Energy and Sustainable Economic Development, Italy</notes><abstract>Agroinfiltration of plant leaves with binary vectors carrying a gene of interest within a plant viral vector is a rapid and efficient method for protein production in plants. Previously, we constructed a self-replicating vector, pA7248AMV, based on the genetic elements of potato virus X (PVX), and have shown that this vector can be used for the expression of recombinant proteins in
. However, this vector is almost 18 kb long and therefore not convenient for genetic manipulation. Furthermore, for efficient expression of the target protein it should be co-agroinfiltrated with an additional binary vector expressing a suppressor of post-transcriptional gene silencing. Here, we improved this expression system by creating the novel pEff vector. Its backbone is about 5 kb shorter than the original vector and it contains an expression cassette for the silencing suppressor, P24, from grapevine leafroll-associated virus-2 alongside PVX genetic elements, thus eliminating the need of co-agroinfiltration. The pEff vector provides green fluorescent protein expression levels of up to 30% of total soluble protein. The novel vector was used for expression of the influenza vaccine candidate, M2eHBc, consisting of an extracellular domain of influenza virus M2 protein (M2e) fused to hepatitis B core antigen. Using the pEff system, M2eHBc was expressed to 5-10% of total soluble protein, several times higher than with original pA7248AMV vector. Plant-produced M2eHBc formed virus-like particles
, as required for its use as a vaccine. The new self-replicating pEff vector could be used for fast and efficient production of various recombinant proteins in plants.</abstract><cop>Switzerland</cop><pub>Frontiers Media S.A</pub><pmid>28293244</pmid><doi>10.3389/fpls.2017.00247</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| title | Efficient Transient Expression of Recombinant Proteins in Plants by the Novel pEff Vector Based on the Genome of Potato Virus X |
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