Expression of the small cysteine-rich protein SCR96 from Phytophthora cactorum in mammalian cells: phytotoxicity and exploitation of its polyclonal antibody.
Identifieur interne : 000213 ( Main/Exploration ); précédent : 000212; suivant : 000214Expression of the small cysteine-rich protein SCR96 from Phytophthora cactorum in mammalian cells: phytotoxicity and exploitation of its polyclonal antibody.
Auteurs : Shen-Xin Huang [République populaire de Chine] ; Zi-Hui Zhang [République populaire de Chine] ; Wang Liu [République populaire de Chine] ; Hang Tao [République populaire de Chine] ; Ye Zhang [République populaire de Chine] ; Ning-Xue Shi [République populaire de Chine] ; Feng Zhu [République populaire de Chine] ; Zhao-Lin Ji [République populaire de Chine] ; Xiao-Ren Chen [République populaire de Chine]Source :
- Biotechnology letters [ 1573-6776 ] ; 2020.
Descripteurs français
- KwdFr :
- Animaux (MeSH), Anticorps (immunologie), Cellules HEK293 (MeSH), Facteurs de virulence (biosynthèse), Facteurs de virulence (génétique), Facteurs de virulence (immunologie), Facteurs de virulence (toxicité), Humains (MeSH), Lapins (MeSH), Lycopersicon esculentum (effets des médicaments et des substances chimiques), Phytophthora (génétique), Phytophthora (métabolisme), Plant (effets des médicaments et des substances chimiques), Protéines fongiques (biosynthèse), Protéines fongiques (génétique), Protéines fongiques (immunologie), Protéines fongiques (toxicité), Protéines recombinantes (biosynthèse), Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Protéines recombinantes (toxicité).
- MESH :
- biosynthèse : Facteurs de virulence, Protéines fongiques, Protéines recombinantes.
- effets des médicaments et des substances chimiques : Lycopersicon esculentum, Plant.
- génétique : Facteurs de virulence, Phytophthora, Protéines fongiques, Protéines recombinantes.
- immunologie : Anticorps, Facteurs de virulence, Protéines fongiques, Protéines recombinantes.
- métabolisme : Phytophthora.
- toxicité : Facteurs de virulence, Protéines fongiques, Protéines recombinantes.
- Animaux, Cellules HEK293, Humains, Lapins.
English descriptors
- KwdEn :
- Animals (MeSH), Antibodies (immunology), Fungal Proteins (biosynthesis), Fungal Proteins (genetics), Fungal Proteins (immunology), Fungal Proteins (toxicity), HEK293 Cells (MeSH), Humans (MeSH), Lycopersicon esculentum (drug effects), Phytophthora (genetics), Phytophthora (metabolism), Rabbits (MeSH), Recombinant Proteins (biosynthesis), Recombinant Proteins (genetics), Recombinant Proteins (immunology), Recombinant Proteins (toxicity), Seedlings (drug effects), Virulence Factors (biosynthesis), Virulence Factors (genetics), Virulence Factors (immunology), Virulence Factors (toxicity).
- MESH :
- chemical , biosynthesis : Fungal Proteins, Recombinant Proteins, Virulence Factors.
- chemical , genetics : Fungal Proteins, Recombinant Proteins, Virulence Factors.
- chemical , immunology : Antibodies, Fungal Proteins, Recombinant Proteins, Virulence Factors.
- chemical , toxicity : Fungal Proteins, Recombinant Proteins, Virulence Factors.
- drug effects : Lycopersicon esculentum, Seedlings.
- genetics : Phytophthora.
- metabolism : Phytophthora.
- Animals, HEK293 Cells, Humans, Rabbits.
Abstract
OBJECTIVE
We aimed to investigate the expression of a novel small cysteine-rich (SCR) effector protein SCR96 from the phytopathogenic oomycete Phytophthora cactorum in mammalian cells, its bioactivity and to exploit its polyclonal antibody.
RESULTS
The gene encoding the SCR effector protein SCR96 was codon-optimized, custom-synthesized, cloned into pcDNA3.1(-) and overexpressed in human embryonic kidney (HEK) 293-6E cells. The recombinant protein SCR96 was prone to aggregation and purified with its monomer to homogeneity with a predicted molecular weight of 8.9 kDa. SCR96 exhibited strong phytotoxic activity on tomato seedlings at 24 h post treatment with 4.2 μg of the purified protein. An anti-SCR96 polyclonal antibody was prepared by immunization of New Zealand white rabbits. The good-titer antibody had a detection sensitivity at 6.25-ng level and could specifically detect the SCR96 protein expressed either in yeast, or in tomato leaves.
CONCLUSIONS
Transient production of the SCR effector protein SCR96 in mammalian cells is reliable, providing sufficient recombinant protein that can be utilized for analysis of its phytotoxic activity and preparation of its polyclonal antibody.
DOI: 10.1007/s10529-019-02754-7
PubMed: 31720976
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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type="wicri:Area/Main/Exploration">000349</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Expression of the small cysteine-rich protein SCR96 from Phytophthora cactorum in mammalian cells: phytotoxicity and exploitation of its polyclonal antibody.</title><author><name sortKey="Huang, Shen Xin" sort="Huang, Shen Xin" uniqKey="Huang S" first="Shen-Xin" last="Huang">Shen-Xin Huang</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author><author><name sortKey="Zhang, Zi Hui" sort="Zhang, Zi Hui" uniqKey="Zhang Z" first="Zi-Hui" last="Zhang">Zi-Hui Zhang</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author><author><name sortKey="Liu, Wang" sort="Liu, Wang" uniqKey="Liu W" first="Wang" last="Liu">Wang Liu</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author><author><name sortKey="Tao, Hang" sort="Tao, Hang" uniqKey="Tao H" first="Hang" last="Tao">Hang Tao</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author><author><name sortKey="Zhang, Ye" sort="Zhang, Ye" uniqKey="Zhang Y" first="Ye" last="Zhang">Ye Zhang</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author><author><name sortKey="Shi, Ning Xue" sort="Shi, Ning Xue" uniqKey="Shi N" first="Ning-Xue" last="Shi">Ning-Xue Shi</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author><author><name sortKey="Zhu, Feng" sort="Zhu, Feng" uniqKey="Zhu F" first="Feng" last="Zhu">Feng Zhu</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author><author><name sortKey="Ji, Zhao Lin" sort="Ji, Zhao Lin" uniqKey="Ji Z" first="Zhao-Lin" last="Ji">Zhao-Lin Ji</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author><author><name sortKey="Chen, Xiao Ren" sort="Chen, Xiao Ren" uniqKey="Chen X" first="Xiao-Ren" last="Chen">Xiao-Ren Chen</name><affiliation wicri:level="1"><nlm:affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China. xrchen@yzu.edu.cn.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China. xrchen@yzu.edu.cn.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province</wicri:regionArea><wicri:noRegion>Jiangsu Province</wicri:noRegion></affiliation></author></analytic><series><title level="j">Biotechnology letters</title><idno type="eISSN">1573-6776</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals (MeSH)</term><term>Antibodies (immunology)</term><term>Fungal Proteins (biosynthesis)</term><term>Fungal Proteins (genetics)</term><term>Fungal Proteins (immunology)</term><term>Fungal Proteins (toxicity)</term><term>HEK293 Cells (MeSH)</term><term>Humans (MeSH)</term><term>Lycopersicon esculentum (drug effects)</term><term>Phytophthora (genetics)</term><term>Phytophthora (metabolism)</term><term>Rabbits (MeSH)</term><term>Recombinant Proteins (biosynthesis)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Recombinant Proteins (toxicity)</term><term>Seedlings (drug effects)</term><term>Virulence Factors (biosynthesis)</term><term>Virulence Factors (genetics)</term><term>Virulence Factors (immunology)</term><term>Virulence Factors (toxicity)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux (MeSH)</term><term>Anticorps (immunologie)</term><term>Cellules HEK293 (MeSH)</term><term>Facteurs de virulence (biosynthèse)</term><term>Facteurs de virulence (génétique)</term><term>Facteurs de virulence (immunologie)</term><term>Facteurs de virulence (toxicité)</term><term>Humains (MeSH)</term><term>Lapins (MeSH)</term><term>Lycopersicon esculentum (effets des médicaments et des substances chimiques)</term><term>Phytophthora (génétique)</term><term>Phytophthora (métabolisme)</term><term>Plant (effets des médicaments et des substances chimiques)</term><term>Protéines fongiques (biosynthèse)</term><term>Protéines fongiques (génétique)</term><term>Protéines fongiques (immunologie)</term><term>Protéines fongiques (toxicité)</term><term>Protéines recombinantes (biosynthèse)</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Protéines recombinantes (toxicité)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Fungal Proteins</term><term>Recombinant Proteins</term><term>Virulence Factors</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Fungal Proteins</term><term>Recombinant Proteins</term><term>Virulence Factors</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies</term><term>Fungal Proteins</term><term>Recombinant Proteins</term><term>Virulence Factors</term></keywords><keywords scheme="MESH" type="chemical" qualifier="toxicity" xml:lang="en"><term>Fungal Proteins</term><term>Recombinant Proteins</term><term>Virulence Factors</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Facteurs de virulence</term><term>Protéines fongiques</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Lycopersicon esculentum</term><term>Seedlings</term></keywords><keywords scheme="MESH" qualifier="effets des médicaments et des substances chimiques" xml:lang="fr"><term>Lycopersicon esculentum</term><term>Plant</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Phytophthora</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Facteurs de virulence</term><term>Phytophthora</term><term>Protéines fongiques</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps</term><term>Facteurs de virulence</term><term>Protéines fongiques</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Phytophthora</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Phytophthora</term></keywords><keywords scheme="MESH" qualifier="toxicité" xml:lang="fr"><term>Facteurs de virulence</term><term>Protéines fongiques</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>HEK293 Cells</term><term>Humans</term><term>Rabbits</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules HEK293</term><term>Humains</term><term>Lapins</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>OBJECTIVE</b></p><p>We aimed to investigate the expression of a novel small cysteine-rich (SCR) effector protein SCR96 from the phytopathogenic oomycete Phytophthora cactorum in mammalian cells, its bioactivity and to exploit its polyclonal antibody.</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>The gene encoding the SCR effector protein SCR96 was codon-optimized, custom-synthesized, cloned into pcDNA3.1(-) and overexpressed in human embryonic kidney (HEK) 293-6E cells. The recombinant protein SCR96 was prone to aggregation and purified with its monomer to homogeneity with a predicted molecular weight of 8.9 kDa. SCR96 exhibited strong phytotoxic activity on tomato seedlings at 24 h post treatment with 4.2 μg of the purified protein. An anti-SCR96 polyclonal antibody was prepared by immunization of New Zealand white rabbits. The good-titer antibody had a detection sensitivity at 6.25-ng level and could specifically detect the SCR96 protein expressed either in yeast, or in tomato leaves.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSIONS</b></p><p>Transient production of the SCR effector protein SCR96 in mammalian cells is reliable, providing sufficient recombinant protein that can be utilized for analysis of its phytotoxic activity and preparation of its polyclonal antibody.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">31720976</PMID><DateCompleted><Year>2020</Year><Month>06</Month><Day>03</Day></DateCompleted><DateRevised><Year>2020</Year><Month>06</Month><Day>03</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1573-6776</ISSN><JournalIssue CitedMedium="Internet"><Volume>42</Volume><Issue>1</Issue><PubDate><Year>2020</Year><Month>Jan</Month></PubDate></JournalIssue><Title>Biotechnology letters</Title><ISOAbbreviation>Biotechnol Lett</ISOAbbreviation></Journal><ArticleTitle>Expression of the small cysteine-rich protein SCR96 from Phytophthora cactorum in mammalian cells: phytotoxicity and exploitation of its polyclonal antibody.</ArticleTitle><Pagination><MedlinePgn>125-133</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1007/s10529-019-02754-7</ELocationID><Abstract><AbstractText Label="OBJECTIVE" NlmCategory="OBJECTIVE">We aimed to investigate the expression of a novel small cysteine-rich (SCR) effector protein SCR96 from the phytopathogenic oomycete Phytophthora cactorum in mammalian cells, its bioactivity and to exploit its polyclonal antibody.</AbstractText><AbstractText Label="RESULTS" NlmCategory="RESULTS">The gene encoding the SCR effector protein SCR96 was codon-optimized, custom-synthesized, cloned into pcDNA3.1(-) and overexpressed in human embryonic kidney (HEK) 293-6E cells. The recombinant protein SCR96 was prone to aggregation and purified with its monomer to homogeneity with a predicted molecular weight of 8.9 kDa. SCR96 exhibited strong phytotoxic activity on tomato seedlings at 24 h post treatment with 4.2 μg of the purified protein. An anti-SCR96 polyclonal antibody was prepared by immunization of New Zealand white rabbits. The good-titer antibody had a detection sensitivity at 6.25-ng level and could specifically detect the SCR96 protein expressed either in yeast, or in tomato leaves.</AbstractText><AbstractText Label="CONCLUSIONS" NlmCategory="CONCLUSIONS">Transient production of the SCR effector protein SCR96 in mammalian cells is reliable, providing sufficient recombinant protein that can be utilized for analysis of its phytotoxic activity and preparation of its polyclonal antibody.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Huang</LastName><ForeName>Shen-Xin</ForeName><Initials>SX</Initials><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Zi-Hui</ForeName><Initials>ZH</Initials><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Wang</ForeName><Initials>W</Initials><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tao</LastName><ForeName>Hang</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Ye</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Shi</LastName><ForeName>Ning-Xue</ForeName><Initials>NX</Initials><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhu</LastName><ForeName>Feng</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Ji</LastName><ForeName>Zhao-Lin</ForeName><Initials>ZL</Initials><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Xiao-Ren</ForeName><Initials>XR</Initials><Identifier Source="ORCID">http://orcid.org/0000-0003-2395-0562</Identifier><AffiliationInfo><Affiliation>College of Horticulture and Plant Protection, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China. xrchen@yzu.edu.cn.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China, Yangzhou University, 48 Wenhui Eastern Road, Yangzhou, 225009, Jiangsu Province, China. xrchen@yzu.edu.cn.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>31671971, 31871907</GrantID><Agency>National Natural Science Foundation of China</Agency><Country></Country></Grant><Grant><GrantID>SJCX19_0895</GrantID><Agency>Postgraduate Research & Practice Innovation Program of Jiangsu Province</Agency><Country></Country></Grant><Grant><GrantID>JRK20180012</GrantID><Agency>the Open Project of Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China</Agency><Country></Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2019</Year><Month>11</Month><Day>12</Day></ArticleDate></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>Biotechnol Lett</MedlineTA><NlmUniqueID>8008051</NlmUniqueID><ISSNLinking>0141-5492</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000906">Antibodies</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D005656">Fungal Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D037521">Virulence Factors</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000906" MajorTopicYN="N">Antibodies</DescriptorName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005656" MajorTopicYN="N">Fungal Proteins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="Y">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName><QualifierName UI="Q000633" MajorTopicYN="Y">toxicity</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D057809" MajorTopicYN="N">HEK293 Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018551" MajorTopicYN="N">Lycopersicon esculentum</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="Y">drug effects</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010838" MajorTopicYN="N">Phytophthora</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011817" MajorTopicYN="N">Rabbits</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName><QualifierName UI="Q000633" MajorTopicYN="N">toxicity</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D036226" MajorTopicYN="N">Seedlings</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D037521" MajorTopicYN="N">Virulence Factors</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="Y">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName><QualifierName UI="Q000633" MajorTopicYN="Y">toxicity</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Ectopic expression</Keyword><Keyword MajorTopicYN="N">Mammalian cell</Keyword><Keyword MajorTopicYN="N">Phytophthora cactorum</Keyword><Keyword MajorTopicYN="N">Phytotoxicity</Keyword><Keyword MajorTopicYN="N">Polyclonal antibody</Keyword><Keyword MajorTopicYN="N">Small cysteine-rich protein</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>08</Month><Day>05</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2019</Year><Month>11</Month><Day>06</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2019</Year><Month>11</Month><Day>14</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>6</Month><Day>4</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2019</Year><Month>11</Month><Day>14</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">31720976</ArticleId><ArticleId IdType="doi">10.1007/s10529-019-02754-7</ArticleId><ArticleId IdType="pii">10.1007/s10529-019-02754-7</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Huang, Shen Xin" sort="Huang, Shen Xin" uniqKey="Huang S" first="Shen-Xin" last="Huang">Shen-Xin Huang</name></noRegion><name sortKey="Chen, Xiao Ren" sort="Chen, Xiao Ren" uniqKey="Chen X" first="Xiao-Ren" last="Chen">Xiao-Ren Chen</name><name sortKey="Chen, Xiao Ren" sort="Chen, Xiao Ren" uniqKey="Chen X" first="Xiao-Ren" last="Chen">Xiao-Ren Chen</name><name sortKey="Huang, Shen Xin" sort="Huang, Shen Xin" uniqKey="Huang S" first="Shen-Xin" last="Huang">Shen-Xin Huang</name><name sortKey="Ji, Zhao Lin" sort="Ji, Zhao Lin" uniqKey="Ji Z" first="Zhao-Lin" last="Ji">Zhao-Lin Ji</name><name sortKey="Ji, Zhao Lin" sort="Ji, Zhao Lin" uniqKey="Ji Z" first="Zhao-Lin" last="Ji">Zhao-Lin Ji</name><name sortKey="Liu, Wang" sort="Liu, Wang" uniqKey="Liu W" first="Wang" last="Liu">Wang Liu</name><name sortKey="Liu, Wang" sort="Liu, Wang" uniqKey="Liu W" first="Wang" last="Liu">Wang Liu</name><name sortKey="Shi, Ning Xue" sort="Shi, Ning Xue" uniqKey="Shi N" first="Ning-Xue" last="Shi">Ning-Xue Shi</name><name sortKey="Shi, Ning Xue" sort="Shi, Ning Xue" uniqKey="Shi N" first="Ning-Xue" last="Shi">Ning-Xue Shi</name><name sortKey="Tao, Hang" sort="Tao, Hang" uniqKey="Tao H" first="Hang" last="Tao">Hang Tao</name><name sortKey="Tao, Hang" sort="Tao, Hang" uniqKey="Tao H" first="Hang" last="Tao">Hang Tao</name><name sortKey="Zhang, Ye" sort="Zhang, Ye" uniqKey="Zhang Y" first="Ye" last="Zhang">Ye Zhang</name><name sortKey="Zhang, Ye" sort="Zhang, Ye" uniqKey="Zhang Y" first="Ye" last="Zhang">Ye Zhang</name><name sortKey="Zhang, Zi Hui" sort="Zhang, Zi Hui" uniqKey="Zhang Z" first="Zi-Hui" last="Zhang">Zi-Hui Zhang</name><name sortKey="Zhang, Zi Hui" sort="Zhang, Zi Hui" uniqKey="Zhang Z" first="Zi-Hui" last="Zhang">Zi-Hui Zhang</name><name sortKey="Zhu, Feng" sort="Zhu, Feng" uniqKey="Zhu F" first="Feng" last="Zhu">Feng Zhu</name><name sortKey="Zhu, Feng" sort="Zhu, Feng" uniqKey="Zhu F" first="Feng" last="Zhu">Feng Zhu</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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