185 lines
17 KiB
XML
185 lines
17 KiB
XML
<document id="D3929DA5338E85EC6622BECC51696622" ID-CLB-Dataset="53911" ID-DOI="10.1016/j.phytochem.2021.112868" ID-GBIF-Dataset="e9409d9e-110c-4801-9730-861bbb65b6bb" ID-ISSN="1873-3700" ID-Zenodo-Dep="8258074" IM.bibliography_approvedBy="felipe" IM.illustrations_approvedBy="felipe" IM.materialsCitations_approvedBy="felipe" IM.metadata_approvedBy="felipe" IM.taxonomicNames_approvedBy="juliana" IM.treatments_approvedBy="juliana" checkinTime="1692304906622" checkinUser="felipe" docAuthor="Zhou, Jiawei, Hu, Tianyuan, Liu, Yuan, Tu, Lichan, Song, Yadi, Lu, Yun, Zhang, Yifeng, Tong, Yuru, Zhao, Yujun, Su, Ping, Wu, Xiaoyi, Huang, Luqi & Gao, Wei" docDate="2021" docId="03F587B4FC0BC8231672FC6CFB6BF84D" docLanguage="en" docName="Phytochemistry.190.112868.pdf" docOrigin="Phytochemistry (112868) 190" docSource="http://dx.doi.org/10.1016/j.phytochem.2021.112868" docStyle="DocumentStyle:F36D69FC8B198FBE91029DF9C24697D3.5:Phytochemistry.2020-.journal_article" docStyleId="F36D69FC8B198FBE91029DF9C24697D3" docStyleName="Phytochemistry.2020-.journal_article" docStyleVersion="5" docTitle="Tripterygium wilfordii subsp. suspension Hook." docType="treatment" docVersion="2" lastPageNumber="8" masterDocId="FFCCFFCCFC0CC8241540FFB5FFCDFFF3" masterDocTitle="Cytochrome P 450 catalyses the 29 - carboxyl group formation of celastrol" masterLastPageNumber="10" masterPageNumber="1" pageNumber="8" updateTime="1692389086640" updateUser="ExternalLinkService">
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<mods:title id="C8F429B4D6C96FF198293452DE69B454">Cytochrome P 450 catalyses the 29 - carboxyl group formation of celastrol</mods:title>
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<mods:namePart id="5CCF50FB2458A0AC05C48B7DA16A1575">Zhou, Jiawei</mods:namePart>
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<mods:affiliation id="DDD71B9132488439F236AA445E6C3FC0">* & Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, China & College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, China & * & School of Traditional Chinese Medicine, Capital Medical University, Beijing, 100069, China</mods:affiliation>
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<mods:namePart id="44E75B742A0118668DF5150B6B111582">Lu, Yun</mods:namePart>
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<mods:namePart id="DE90BAF21AF3FF4C947DCE12C673C3B5">Wu, Xiaoyi</mods:namePart>
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<mods:namePart id="F97578616125321FCD2F6BF04B5A9F6D">Huang, Luqi</mods:namePart>
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<mods:namePart id="3DD0A1442DA81C82094B850D1D8F0FB2">Gao, Wei</mods:namePart>
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<mods:title id="32F6C946D8BF2B6B9AB9C8AD4C61164C">Phytochemistry</mods:title>
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<mods:date id="BBB0F333CD7656BD479B35F27C218D9B">2021</mods:date>
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<treatment id="03F587B4FC0BC8231672FC6CFB6BF84D" ID-DOI="http://doi.org/10.5281/zenodo.8264301" ID-Zenodo-Dep="8264301" LSID="urn:lsid:plazi:treatment:03F587B4FC0BC8231672FC6CFB6BF84D" httpUri="http://treatment.plazi.org/id/03F587B4FC0BC8231672FC6CFB6BF84D" lastPageNumber="8" pageId="7" pageNumber="8">
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4.7. Gene expression level network analysis for celastrol in
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<taxonomicName id="4C5C4D21FC0BC823100DFC6FFA7CFC1F" ID-CoL="7D6YZ" ID-ENA="458696" authority="Hook." authorityName="Hook." box="[1357,1457,985,1005]" class="Magnoliopsida" family="Celastraceae" genus="Tripterygium" kingdom="Plantae" order="Celastrales" pageId="7" pageNumber="8" phylum="Tracheophyta" rank="species" species="wilfordii">T. wilfordii</taxonomicName>
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<paragraph id="8BE336A2FC0BC8231611FBA4FC21FAC8" blockId="7.[818,1487,1041,1339]" pageId="7" pageNumber="8">
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The gene expression levels of all CYP450s and
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<emphasis id="B928EAB0FC0BC8231052FBA4FAE0FBD7" bold="true" box="[1298,1325,1041,1060]" italics="true" pageId="7" pageNumber="8">Tw</emphasis>
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OSCs in different
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<taxonomicName id="4C5C4D21FC0BC8231672FB98FC58FBB3" box="[818,917,1069,1088]" class="Magnoliopsida" family="Celastraceae" genus="Tripterygium" kingdom="Plantae" order="Celastrales" pageId="7" pageNumber="8" phylum="Tracheophyta" rank="species" species="wilfordii">
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<emphasis id="B928EAB0FC0BC8231672FB98FC58FBB3" bold="true" box="[818,917,1069,1088]" italics="true" pageId="7" pageNumber="8">T. wilfordii</emphasis>
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</taxonomicName>
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tissues were chosen for network regulation analysis associated with celastrol levels in different
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<taxonomicName id="4C5C4D21FC0BC82311D0FBFCFB3FFBAF" box="[1168,1266,1097,1116]" class="Magnoliopsida" family="Celastraceae" genus="Tripterygium" kingdom="Plantae" order="Celastrales" pageId="7" pageNumber="8" phylum="Tracheophyta" rank="species" species="wilfordii">
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<emphasis id="B928EAB0FC0BC82311D0FBFCFB3FFBAF" bold="true" box="[1168,1266,1097,1116]" italics="true" pageId="7" pageNumber="8">T. wilfordii</emphasis>
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</taxonomicName>
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tissues. Celastrol levels in different tissues were detected as previously described (
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<bibRefCitation id="EFCD4B53FC0BC8231022FBD0FCAFFB67" author="Zhou, J. & Hu, T. & Gao, L. & Su, P. & Zhang, Y. & Zhao, Y. & Chen, S. & Tu, L. & Song, Y. & Wang, X. & Huang, L. & Gao, W." pageId="7" pageNumber="8" pagination="722 - 735" refId="ref11593" refString="Zhou, J., Hu, T., Gao, L., Su, P., Zhang, Y., Zhao, Y., Chen, S., Tu, L., Song, Y., Wang, X., Huang, L., Gao, W., 2019. Friedelane-type triterpene cyclase in celastrol biosynthesis from Tripterygium wilfordii and its application for triterpenes biosynthesis in yeast. New Phytol. 223, 722 - 735. https: // doi. org / 10.1111 / nph. 15809." type="journal article" year="2019">Zhou et al., 2019</bibRefCitation>
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). The celastrol level and gene expression data were normalized separately, and correlation network analysis was conducted with Cytoscape software (version 3.6.1). The Pearson correlation coefficient was calculated via the PCC method on the R platform between each set of variables (either metabolite or gene) across the profiles, and significant positive correlations with a p-value
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<emphasis id="B928EAB0FC0BC82311C8FAB8FB55FAD3" box="[1160,1176,1293,1312]" italics="true" pageId="7" pageNumber="8"><</emphasis>
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0.05 were detected between the genes and celastrol.
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</paragraph>
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<paragraph id="8BE336A2FC0BC8231672FAD4FBC0FA7C" blockId="7.[818,1409,1376,1396]" lastBlockId="7.[818,1037,1404,1424]" pageId="7" pageNumber="8">
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<emphasis id="B928EAB0FC0BC8231672FAD4FA4CFA87" bold="true" box="[818,1409,1376,1396]" italics="true" pageId="7" pageNumber="8">
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4.8. RNAi knockdown and overexpression of TwCYP712K
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<quantity id="4CA49B47FC0BC8231006FAD4FAA5FA87" box="[1350,1384,1377,1396]" metricMagnitude="-2" metricUnit="m" metricValue="2.54" pageId="7" pageNumber="8" unit="in" value="1.0">1 in</quantity>
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<taxonomicName id="4C5C4D21FC0BC823102FFAD4FCB1FA7C" class="Magnoliopsida" family="Celastraceae" genus="Tripterygium" kingdom="Plantae" order="Celastrales" pageId="7" pageNumber="8" phylum="Tracheophyta" rank="species" species="wilfordii">
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T.
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<emphasis id="B928EAB0FC0BC8231672FAC9FBC0FA7C" bold="true" box="[818,1037,1404,1424]" italics="true" pageId="7" pageNumber="8">wilfordii suspension cells</emphasis>
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</taxonomicName>
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</emphasis>
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</paragraph>
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<paragraph id="8BE336A2FC0BC8231611FA00FB6BF84D" blockId="7.[818,1488,1460,1982]" pageId="7" pageNumber="8">
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For the RNAi knockdown of
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<emphasis id="B928EAB0FC0BC8231131FA01FB3DFA34" bold="true" box="[1137,1264,1460,1479]" italics="true" pageId="7" pageNumber="8">TwCYP712K1</emphasis>
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, the least homologous region of
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<emphasis id="B928EAB0FC0BC82316D3FA65FBDFFA10" bold="true" box="[915,1042,1488,1507]" italics="true" pageId="7" pageNumber="8">TwCYP712K1</emphasis>
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was selected based on cDNA alignment, and specific primers were then designed according to the sequence of the region (Supplementary Table S4).
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<collectionCode id="ED4DAE67FC0BC8231137F9BDFB48F9E8" box="[1143,1157,1544,1563]" country="USA" lsid="urn:lsid:biocol.org:col:15406" name="Harvard University - Arnold Arboretum" pageId="7" pageNumber="8" type="Herbarium">A</collectionCode>
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specific 300–500 bp fragment was amplified and inserted into the pK7GWIWG2D vector (Invitrogen) using the Gateway cloning system (Invitrogen). For the overexpression of
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<emphasis id="B928EAB0FC0BC8231672F9E9FC7CF99C" bold="true" box="[818,945,1628,1647]" italics="true" pageId="7" pageNumber="8">TwCYP712K1</emphasis>
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, the full-length sequence of
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<emphasis id="B928EAB0FC0BC8231184F9E9FA8EF99C" bold="true" box="[1220,1347,1628,1647]" italics="true" pageId="7" pageNumber="8">TwCYP712K1</emphasis>
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was amplified and subcloned into the pH7WG2D vector (Invitrogen). The resulting vectors were separately transformed into
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<taxonomicName id="4C5C4D21FC0BC8231188F921FA50F954" box="[1224,1437,1683,1703]" class="Magnoliopsida" family="Celastraceae" genus="Tripterygium" kingdom="Plantae" order="Celastrales" pageId="7" pageNumber="8" phylum="Tracheophyta" rank="subSpecies" species="wilfordii" subSpecies="suspension">
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<emphasis id="B928EAB0FC0BC8231188F921FAE0F955" bold="true" box="[1224,1325,1683,1703]" italics="true" pageId="7" pageNumber="8">T. wilfordii</emphasis>
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suspension
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</taxonomicName>
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cells based on particle bombardment-mediated transformation. The detailed processes of suspension cell preparation and transformation were the same as in a previous report (
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<bibRefCitation id="EFCD4B53FC0BC8231103F952FB12F909" author="Zhao, Y. & Zhang, Y. & Su, P. & Yang, J. & Huang, L. & Gao, W." box="[1091,1247,1767,1787]" pageId="7" pageNumber="8" pagination="2221" refId="ref11523" refString="Zhao, Y., Zhang, Y., Su, P., Yang, J., Huang, L., Gao, W., 2017. Genetic transformation system for woody plant Tripterygium wilfordii and its application to product natural celastrol. Front. Plant Sci. 8, 2221. https: // doi. org / 10.3389 / fpls. 2017.02221." type="journal article" year="2017">Zhao et al., 2017</bibRefCitation>
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||
). The pK7GWIWG2D and pH7WG2D vectors were separately transformed into
|
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<taxonomicName id="4C5C4D21FC0BC823107BF8B6FCB6F8C1" class="Magnoliopsida" family="Celastraceae" genus="Tripterygium" kingdom="Plantae" order="Celastrales" pageId="7" pageNumber="8" phylum="Tracheophyta" rank="subSpecies" species="wilfordii" subSpecies="suspension">
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<emphasis id="B928EAB0FC0BC823107BF8B6FA6CF8E5" bold="true" box="[1339,1441,1795,1814]" italics="true" pageId="7" pageNumber="8">T. wilfordii</emphasis>
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suspension
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</taxonomicName>
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cells as controls, and at least three biological duplicates were included for all samples. The cell suspensions were cultured for another 7 days after transformation, and real-time PCR and UPLC analyses were conducted to measure gene expression and celastrol levels, respectively. The detailed process of UPLC analysis was the same as that described in our previous report (
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<bibRefCitation id="EFCD4B53FC0BC82316B7F81EFB54F84D" author="Zhou, J. & Hu, T. & Gao, L. & Su, P. & Zhang, Y. & Zhao, Y. & Chen, S. & Tu, L. & Song, Y. & Wang, X. & Huang, L. & Gao, W." box="[1015,1177,1963,1982]" pageId="7" pageNumber="8" pagination="722 - 735" refId="ref11593" refString="Zhou, J., Hu, T., Gao, L., Su, P., Zhang, Y., Zhao, Y., Chen, S., Tu, L., Song, Y., Wang, X., Huang, L., Gao, W., 2019. Friedelane-type triterpene cyclase in celastrol biosynthesis from Tripterygium wilfordii and its application for triterpenes biosynthesis in yeast. New Phytol. 223, 722 - 735. https: // doi. org / 10.1111 / nph. 15809." type="journal article" year="2019">Zhou et al., 2019</bibRefCitation>
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).
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</paragraph>
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</subSubSection>
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</treatment>
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</document> |