Fusion-PCR generates attL recombination site adaptors and allows Rapid One-Step Gateway (ROG) cloning.
Identifieur interne : 000038 ( Main/Exploration ); précédent : 000037; suivant : 000039Fusion-PCR generates attL recombination site adaptors and allows Rapid One-Step Gateway (ROG) cloning.
Auteurs : Libing Liao [République populaire de Chine] ; Lu Yang [République populaire de Chine] ; Yanxia Xu [République populaire de Chine] ; Xiaoli Li [République populaire de Chine] ; Guangxuan Tan [République populaire de Chine] ; Beibei Fu [République populaire de Chine] ; Kelei Duan [République populaire de Chine] ; Zhiqiang Li [République populaire de Chine] ; Deshui Yu [République populaire de Chine] ; Chengwei Li [République populaire de Chine]Source :
- Biochimie [ 1638-6183 ] ; 2020.
Abstract
Gateway recombination-based cloning, which eliminates the use of restriction endonucleases and ligase, has been widely used for the construction of high-throughput (HTP) vectors. However, this approach is very expensive and its two-stage reaction process is laborious and time consuming. Therefore, we developed a Gateway cloning method that uses fusion-PCR to generate attL recombination site adaptors, and the PCR products, which can be directly cloned into destination vectors, giving rise to Rapid One-Step Gateway (ROG) Cloning. 100% of cloning efficiencies were obtained by this ROG method. This method has no BP reaction/entry clone step, thus halving the cost and time consumed. Overall, this work provides a highly efficient, rapid, low-cost method for directional recombination cloning.
DOI: 10.1016/j.biochi.2020.04.002
PubMed: 32325113
Affiliations:
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Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Li, Xiaoli" sort="Li, Xiaoli" uniqKey="Li X" first="Xiaoli" last="Li">Xiaoli Li</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; 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Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Duan, Kelei" sort="Duan, Kelei" uniqKey="Duan K" first="Kelei" last="Duan">Kelei Duan</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Li, Zhiqiang" sort="Li, Zhiqiang" uniqKey="Li Z" first="Zhiqiang" last="Li">Zhiqiang Li</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Yu, Deshui" sort="Yu, Deshui" uniqKey="Yu D" first="Deshui" last="Yu">Deshui Yu</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Lushan Biotanical Garden, Chinese Academy of Science, Jiujiang, 332900, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China. 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Lushan Biotanical Garden, Chinese Academy of Science, Jiujiang, 332900, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Lushan Biotanical Garden, Chinese Academy of Science, Jiujiang, 332900, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Lu" sort="Yang, Lu" uniqKey="Yang L" first="Lu" last="Yang">Lu Yang</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Xu, Yanxia" sort="Xu, Yanxia" uniqKey="Xu Y" first="Yanxia" last="Xu">Yanxia Xu</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Li, Xiaoli" sort="Li, Xiaoli" uniqKey="Li X" first="Xiaoli" last="Li">Xiaoli Li</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Tan, Guangxuan" sort="Tan, Guangxuan" uniqKey="Tan G" first="Guangxuan" last="Tan">Guangxuan Tan</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Fu, Beibei" sort="Fu, Beibei" uniqKey="Fu B" first="Beibei" last="Fu">Beibei Fu</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Duan, Kelei" sort="Duan, Kelei" uniqKey="Duan K" first="Kelei" last="Duan">Kelei Duan</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Li, Zhiqiang" sort="Li, Zhiqiang" uniqKey="Li Z" first="Zhiqiang" last="Li">Zhiqiang Li</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Yu, Deshui" sort="Yu, Deshui" uniqKey="Yu D" first="Deshui" last="Yu">Deshui Yu</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Lushan Biotanical Garden, Chinese Academy of Science, Jiujiang, 332900, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China. Electronic address: ds-yu@163.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Lushan Biotanical Garden, Chinese Academy of Science, Jiujiang, 332900, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001</wicri:regionArea><wicri:noRegion>466001</wicri:noRegion></affiliation></author><author><name sortKey="Li, Chengwei" sort="Li, Chengwei" uniqKey="Li C" first="Chengwei" last="Li">Chengwei Li</name><affiliation wicri:level="1"><nlm:affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang, 453003, China. Electronic address: lichengweihist@163.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang, 453003</wicri:regionArea><wicri:noRegion>453003</wicri:noRegion></affiliation></author></analytic><series><title level="j">Biochimie</title><idno type="eISSN">1638-6183</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Gateway recombination-based cloning, which eliminates the use of restriction endonucleases and ligase, has been widely used for the construction of high-throughput (HTP) vectors. However, this approach is very expensive and its two-stage reaction process is laborious and time consuming. Therefore, we developed a Gateway cloning method that uses fusion-PCR to generate attL recombination site adaptors, and the PCR products, which can be directly cloned into destination vectors, giving rise to Rapid One-Step Gateway (ROG) Cloning. 100% of cloning efficiencies were obtained by this ROG method. This method has no BP reaction/entry clone step, thus halving the cost and time consumed. Overall, this work provides a highly efficient, rapid, low-cost method for directional recombination cloning.</div></front></TEI><pubmed><MedlineCitation Status="In-Process" Owner="NLM"><PMID Version="1">32325113</PMID><DateRevised><Year>2020</Year><Month>06</Month><Day>08</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1638-6183</ISSN><JournalIssue CitedMedium="Internet"><Volume>174</Volume><PubDate><Year>2020</Year><Month>Jul</Month></PubDate></JournalIssue><Title>Biochimie</Title><ISOAbbreviation>Biochimie</ISOAbbreviation></Journal><ArticleTitle>Fusion-PCR generates attL recombination site adaptors and allows Rapid One-Step Gateway (ROG) cloning.</ArticleTitle><Pagination><MedlinePgn>69-73</MedlinePgn></Pagination><ELocationID EIdType="pii" ValidYN="Y">S0300-9084(20)30072-9</ELocationID><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.biochi.2020.04.002</ELocationID><Abstract><AbstractText>Gateway recombination-based cloning, which eliminates the use of restriction endonucleases and ligase, has been widely used for the construction of high-throughput (HTP) vectors. However, this approach is very expensive and its two-stage reaction process is laborious and time consuming. Therefore, we developed a Gateway cloning method that uses fusion-PCR to generate attL recombination site adaptors, and the PCR products, which can be directly cloned into destination vectors, giving rise to Rapid One-Step Gateway (ROG) Cloning. 100% of cloning efficiencies were obtained by this ROG method. This method has no BP reaction/entry clone step, thus halving the cost and time consumed. Overall, this work provides a highly efficient, rapid, low-cost method for directional recombination cloning.</AbstractText><CopyrightInformation>Copyright © 2020 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Liao</LastName><ForeName>Libing</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Lushan Biotanical Garden, Chinese Academy of Science, Jiujiang, 332900, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Lu</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Xu</LastName><ForeName>Yanxia</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Xiaoli</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tan</LastName><ForeName>Guangxuan</ForeName><Initials>G</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fu</LastName><ForeName>Beibei</ForeName><Initials>B</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Duan</LastName><ForeName>Kelei</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Zhiqiang</ForeName><Initials>Z</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, 466001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yu</LastName><ForeName>Deshui</ForeName><Initials>D</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Lushan Biotanical Garden, Chinese Academy of Science, Jiujiang, 332900, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China. Electronic address: ds-yu@163.com.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Chengwei</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, 466001, China; Henan Key Laboratory of Crop Molecular Breeding & Bioreactor, Zhoukou, 466001, China; Henan Engineering Research Center of Crop Genome Editing, Henan Institute of Science and Technology, Xinxiang, 453003, China. Electronic address: lichengweihist@163.com.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>04</Month><Day>20</Day></ArticleDate></Article><MedlineJournalInfo><Country>France</Country><MedlineTA>Biochimie</MedlineTA><NlmUniqueID>1264604</NlmUniqueID><ISSNLinking>0300-9084</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Agrobacterium-mediated transient expression assay</Keyword><Keyword MajorTopicYN="N">Fusion-PCR</Keyword><Keyword MajorTopicYN="N">Rapid One-Step Gateway (ROG) cloning</Keyword></KeywordList><CoiStatement>Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.</CoiStatement></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2020</Year><Month>01</Month><Day>13</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2020</Year><Month>03</Month><Day>31</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2020</Year><Month>04</Month><Day>07</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>4</Month><Day>24</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>4</Month><Day>24</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>4</Month><Day>24</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">32325113</ArticleId><ArticleId IdType="pii">S0300-9084(20)30072-9</ArticleId><ArticleId IdType="doi">10.1016/j.biochi.2020.04.002</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="Liao, Libing" sort="Liao, Libing" uniqKey="Liao L" first="Libing" last="Liao">Libing Liao</name></noRegion><name sortKey="Duan, Kelei" sort="Duan, Kelei" uniqKey="Duan K" first="Kelei" last="Duan">Kelei Duan</name><name sortKey="Fu, Beibei" sort="Fu, Beibei" uniqKey="Fu B" first="Beibei" last="Fu">Beibei Fu</name><name sortKey="Li, Chengwei" sort="Li, Chengwei" uniqKey="Li C" first="Chengwei" last="Li">Chengwei Li</name><name sortKey="Li, Xiaoli" sort="Li, Xiaoli" uniqKey="Li X" first="Xiaoli" last="Li">Xiaoli Li</name><name sortKey="Li, Zhiqiang" sort="Li, Zhiqiang" uniqKey="Li Z" first="Zhiqiang" last="Li">Zhiqiang Li</name><name sortKey="Tan, Guangxuan" sort="Tan, Guangxuan" uniqKey="Tan G" first="Guangxuan" last="Tan">Guangxuan Tan</name><name sortKey="Xu, Yanxia" sort="Xu, Yanxia" uniqKey="Xu Y" first="Yanxia" last="Xu">Yanxia Xu</name><name sortKey="Yang, Lu" sort="Yang, Lu" uniqKey="Yang L" first="Lu" last="Yang">Lu Yang</name><name sortKey="Yu, Deshui" sort="Yu, Deshui" uniqKey="Yu D" first="Deshui" last="Yu">Deshui Yu</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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