[A hydroponic cultivation system for rapid high-yield transient protein expression in Nicotiana plants under laboratory conditions].
Identifieur interne : 000493 ( Main/Exploration ); précédent : 000492; suivant : 000494[A hydroponic cultivation system for rapid high-yield transient protein expression in Nicotiana plants under laboratory conditions].
Auteurs : Qianzhen Mo [République populaire de Chine] ; Rongjia Mai ; Zhixiao Yang ; Minfang Chen ; Tiezhao Yang ; Huafang Lai ; Peiliang Yang ; Qiang Chen ; Xiaohong ZhouSource :
- Nan fang yi ke da xue xue bao = Journal of Southern Medical University [ 1673-4254 ] ; 2012.
Descripteurs français
- KwdFr :
- Culture hydroponique (méthodes), Protéines recombinantes (biosynthèse), Protéines recombinantes (génétique), Protéines à fluorescence verte (biosynthèse), Protéines à fluorescence verte (génétique), Régulation de l'expression des gènes végétaux (MeSH), Tabac (croissance et développement), Tabac (génétique), Vecteurs génétiques (MeSH), Végétaux génétiquement modifiés (génétique).
- MESH :
- biosynthèse : Protéines recombinantes, Protéines à fluorescence verte.
- croissance et développement : Tabac.
- génétique : Protéines recombinantes, Protéines à fluorescence verte, Tabac, Végétaux génétiquement modifiés.
- méthodes : Culture hydroponique.
- Régulation de l'expression des gènes végétaux, Vecteurs génétiques.
English descriptors
- KwdEn :
- Gene Expression Regulation, Plant (MeSH), Genetic Vectors (MeSH), Green Fluorescent Proteins (biosynthesis), Green Fluorescent Proteins (genetics), Hydroponics (methods), Plants, Genetically Modified (genetics), Recombinant Proteins (biosynthesis), Recombinant Proteins (genetics), Tobacco (genetics), Tobacco (growth & development).
- MESH :
- chemical , biosynthesis : Green Fluorescent Proteins, Recombinant Proteins.
- chemical , genetics : Green Fluorescent Proteins, Recombinant Proteins.
- genetics : Plants, Genetically Modified, Tobacco.
- growth & development : Tobacco.
- methods : Hydroponics.
- Gene Expression Regulation, Plant, Genetic Vectors.
Abstract
OBJECTIVE
To develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions.
METHODS
To establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA.
RESULTS
Our data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration.
CONCLUSION
We have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.
PubMed: 22699052
Affiliations:
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sortKey="Chen, Minfang" sort="Chen, Minfang" uniqKey="Chen M" first="Minfang" last="Chen">Minfang Chen</name></author><author><name sortKey="Yang, Tiezhao" sort="Yang, Tiezhao" uniqKey="Yang T" first="Tiezhao" last="Yang">Tiezhao Yang</name></author><author><name sortKey="Lai, Huafang" sort="Lai, Huafang" uniqKey="Lai H" first="Huafang" last="Lai">Huafang Lai</name></author><author><name sortKey="Yang, Peiliang" sort="Yang, Peiliang" uniqKey="Yang P" first="Peiliang" last="Yang">Peiliang Yang</name></author><author><name sortKey="Chen, Qiang" sort="Chen, Qiang" uniqKey="Chen Q" first="Qiang" last="Chen">Qiang Chen</name></author><author><name sortKey="Zhou, Xiaohong" sort="Zhou, Xiaohong" uniqKey="Zhou X" first="Xiaohong" last="Zhou">Xiaohong Zhou</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2012">2012</date><idno type="RBID">pubmed:22699052</idno><idno type="pmid">22699052</idno><idno type="wicri:Area/Main/Corpus">000509</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000509</idno><idno type="wicri:Area/Main/Curation">000509</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000509</idno><idno type="wicri:Area/Main/Exploration">000509</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">[A hydroponic cultivation system for rapid high-yield transient protein expression in Nicotiana plants under laboratory conditions].</title><author><name sortKey="Mo, Qianzhen" sort="Mo, Qianzhen" uniqKey="Mo Q" first="Qianzhen" last="Mo">Qianzhen Mo</name><affiliation wicri:level="3"><nlm:affiliation>Southern Medical University, Guangzhou, China. qianzhen.love@163.com</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Southern Medical University, Guangzhou</wicri:regionArea><placeName><settlement type="city">Jiangmen</settlement><region type="province">Guangdong</region></placeName></affiliation></author><author><name sortKey="Mai, Rongjia" sort="Mai, Rongjia" uniqKey="Mai R" first="Rongjia" last="Mai">Rongjia Mai</name></author><author><name sortKey="Yang, Zhixiao" sort="Yang, Zhixiao" uniqKey="Yang Z" first="Zhixiao" last="Yang">Zhixiao Yang</name></author><author><name sortKey="Chen, Minfang" sort="Chen, Minfang" uniqKey="Chen M" first="Minfang" last="Chen">Minfang Chen</name></author><author><name sortKey="Yang, Tiezhao" sort="Yang, Tiezhao" uniqKey="Yang T" first="Tiezhao" last="Yang">Tiezhao Yang</name></author><author><name sortKey="Lai, Huafang" sort="Lai, Huafang" uniqKey="Lai H" first="Huafang" last="Lai">Huafang Lai</name></author><author><name sortKey="Yang, Peiliang" sort="Yang, Peiliang" uniqKey="Yang P" first="Peiliang" last="Yang">Peiliang Yang</name></author><author><name sortKey="Chen, Qiang" sort="Chen, Qiang" uniqKey="Chen Q" first="Qiang" last="Chen">Qiang Chen</name></author><author><name sortKey="Zhou, Xiaohong" sort="Zhou, Xiaohong" uniqKey="Zhou X" first="Xiaohong" last="Zhou">Xiaohong Zhou</name></author></analytic><series><title level="j">Nan fang yi ke da xue xue bao = Journal of Southern Medical University</title><idno type="ISSN">1673-4254</idno><imprint><date when="2012" type="published">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Gene Expression Regulation, Plant (MeSH)</term><term>Genetic Vectors (MeSH)</term><term>Green Fluorescent Proteins (biosynthesis)</term><term>Green Fluorescent Proteins (genetics)</term><term>Hydroponics (methods)</term><term>Plants, Genetically Modified (genetics)</term><term>Recombinant Proteins (biosynthesis)</term><term>Recombinant Proteins (genetics)</term><term>Tobacco (genetics)</term><term>Tobacco (growth & development)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Culture hydroponique (méthodes)</term><term>Protéines recombinantes (biosynthèse)</term><term>Protéines recombinantes (génétique)</term><term>Protéines à fluorescence verte (biosynthèse)</term><term>Protéines à fluorescence verte (génétique)</term><term>Régulation de l'expression des gènes végétaux (MeSH)</term><term>Tabac (croissance et développement)</term><term>Tabac (génétique)</term><term>Vecteurs génétiques (MeSH)</term><term>Végétaux génétiquement modifiés (génétique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Green Fluorescent Proteins</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Green Fluorescent Proteins</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Protéines recombinantes</term><term>Protéines à fluorescence verte</term></keywords><keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Tabac</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Plants, Genetically Modified</term><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>Tobacco</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines recombinantes</term><term>Protéines à fluorescence verte</term><term>Tabac</term><term>Végétaux génétiquement modifiés</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Hydroponics</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Culture hydroponique</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Gene Expression Regulation, Plant</term><term>Genetic Vectors</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Régulation de l'expression des gènes végétaux</term><term>Vecteurs génétiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>OBJECTIVE</b></p><p>To develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions.</p></div><div type="abstract" xml:lang="en"><p><b>METHODS</b></p><p>To establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA.</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>Our data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSION</b></p><p>We have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">22699052</PMID><DateCompleted><Year>2013</Year><Month>05</Month><Day>23</Day></DateCompleted><DateRevised><Year>2012</Year><Month>06</Month><Day>15</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">1673-4254</ISSN><JournalIssue CitedMedium="Print"><Volume>32</Volume><Issue>6</Issue><PubDate><Year>2012</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Nan fang yi ke da xue xue bao = Journal of Southern Medical University</Title><ISOAbbreviation>Nan Fang Yi Ke Da Xue Xue Bao</ISOAbbreviation></Journal><ArticleTitle>[A hydroponic cultivation system for rapid high-yield transient protein expression in Nicotiana plants under laboratory conditions].</ArticleTitle><Pagination><MedlinePgn>772-7</MedlinePgn></Pagination><Abstract><AbstractText Label="OBJECTIVE" NlmCategory="OBJECTIVE">To develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions.</AbstractText><AbstractText Label="METHODS" NlmCategory="METHODS">To establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA.</AbstractText><AbstractText Label="RESULTS" NlmCategory="RESULTS">Our data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration.</AbstractText><AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">We have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Mo</LastName><ForeName>Qianzhen</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>Southern Medical University, Guangzhou, China. qianzhen.love@163.com</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Mai</LastName><ForeName>Rongjia</ForeName><Initials>R</Initials></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Zhixiao</ForeName><Initials>Z</Initials></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Minfang</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Tiezhao</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Lai</LastName><ForeName>Huafang</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Peiliang</ForeName><Initials>P</Initials></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Qiang</ForeName><Initials>Q</Initials></Author><Author ValidYN="Y"><LastName>Zhou</LastName><ForeName>Xiaohong</ForeName><Initials>X</Initials></Author></AuthorList><Language>chi</Language><PublicationTypeList><PublicationType UI="D004740">English Abstract</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>China</Country><MedlineTA>Nan Fang Yi Ke Da Xue Xue Bao</MedlineTA><NlmUniqueID>101266132</NlmUniqueID><ISSNLinking>1673-4254</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>147336-22-9</RegistryNumber><NameOfSubstance UI="D049452">Green Fluorescent Proteins</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D018506" MajorTopicYN="N">Gene Expression Regulation, Plant</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005822" MajorTopicYN="N">Genetic Vectors</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D049452" MajorTopicYN="N">Green Fluorescent Proteins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D018527" MajorTopicYN="N">Hydroponics</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D030821" MajorTopicYN="N">Plants, Genetically Modified</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="Y">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014026" MajorTopicYN="N">Tobacco</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000254" MajorTopicYN="N">growth & development</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="entrez"><Year>2012</Year><Month>6</Month><Day>16</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2012</Year><Month>6</Month><Day>16</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2013</Year><Month>5</Month><Day>25</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">22699052</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Guangdong</li></region><settlement><li>Jiangmen</li></settlement></list><tree><noCountry><name sortKey="Chen, Minfang" sort="Chen, Minfang" uniqKey="Chen M" first="Minfang" last="Chen">Minfang Chen</name><name sortKey="Chen, Qiang" sort="Chen, Qiang" uniqKey="Chen Q" first="Qiang" last="Chen">Qiang Chen</name><name sortKey="Lai, Huafang" sort="Lai, Huafang" uniqKey="Lai H" first="Huafang" last="Lai">Huafang Lai</name><name sortKey="Mai, Rongjia" sort="Mai, Rongjia" uniqKey="Mai R" first="Rongjia" last="Mai">Rongjia Mai</name><name sortKey="Yang, Peiliang" sort="Yang, Peiliang" uniqKey="Yang P" first="Peiliang" last="Yang">Peiliang Yang</name><name sortKey="Yang, Tiezhao" sort="Yang, Tiezhao" uniqKey="Yang T" first="Tiezhao" last="Yang">Tiezhao Yang</name><name sortKey="Yang, Zhixiao" sort="Yang, Zhixiao" uniqKey="Yang Z" first="Zhixiao" last="Yang">Zhixiao Yang</name><name sortKey="Zhou, Xiaohong" sort="Zhou, Xiaohong" uniqKey="Zhou X" first="Xiaohong" last="Zhou">Xiaohong Zhou</name></noCountry><country name="République populaire de Chine"><region name="Guangdong"><name sortKey="Mo, Qianzhen" sort="Mo, Qianzhen" uniqKey="Mo Q" first="Qianzhen" last="Mo">Qianzhen Mo</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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