130 lines
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130 lines
13 KiB
XML
<document id="F01B7446D82731C9BCA4E9B367FA5700" ID-DOI="10.1016/j.phytochem.2020.112595" ID-ISSN="1873-3700" ID-Zenodo-Dep="8291489" IM.bibliography_approvedBy="juliana" IM.illustrations_approvedBy="juliana" IM.materialsCitations_approvedBy="felipe" IM.metadata_approvedBy="juliana" IM.tables_approvedBy="juliana" IM.taxonomicNames_approvedBy="juliana" IM.treatments_approvedBy="juliana" checkinTime="1693245633458" checkinUser="felipe" docAuthor="Cruz-Silva, Ilana, Gozzo, Andrezza Justino, Nunes, Viviane Abreu, Tanaka, Aparecida Sadae & Araujo, Mariana da Silva" docDate="2021" docId="03FC87C47220FF89FB03107AFA78FD20" docLanguage="en" docName="Phytochemistry.182.112595.pdf" docOrigin="Phytochemistry (112595) 182" docSource="http://dx.doi.org/10.1016/j.phytochem.2020.112595" docStyle="DocumentStyle:F36D69FC8B198FBE91029DF9C24697D3.5:Phytochemistry.2020-.journal_article" docStyleId="F36D69FC8B198FBE91029DF9C24697D3" docStyleName="Phytochemistry.2020-.journal_article" docStyleVersion="5" docTitle="Caesalpinia echinata" docType="treatment" docVersion="1" lastPageNumber="6" masterDocId="FFC5FFBC7225FF8CFB671621FFA8FFA1" masterDocTitle="Bioengineering of an elastase inhibitor from Caesalpinia echinata (Brazil wood) seeds" masterLastPageNumber="10" masterPageNumber="1" pageNumber="6" updateTime="1693403065341" updateUser="juliana">
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<mods:title id="9A3E3E337527501F3CDBB911809E5F2F">Bioengineering of an elastase inhibitor from Caesalpinia echinata (Brazil wood) seeds</mods:title>
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<mods:namePart id="CEEF04B1B0305BAC29E784A90600A38D">Cruz-Silva, Ilana</mods:namePart>
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<mods:affiliation id="C14DE2E49798EDAB961AA4B76588AF08">Department of Biochemistry, Universidade Federal de S ˜ ao Paulo, Rua Trˆes de Maio, 100, 04044 - 020, S ˜ ao Paulo, SP, Brazil & Division of Dermatology, Hospital Sírio Libanˆes, Rua Professor Daher Cutait, 69, 01308 - 060, S ˜ ao Paulo, SP, Brazil</mods:affiliation>
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<mods:namePart id="618678EEA38A24265CC4550F20D5C7EF">Gozzo, Andrezza Justino</mods:namePart>
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<mods:affiliation id="A0497764DDC2DCD1062D1605E39F6C0D">** & Institute of Marine Sciences, Universidade Federal de S ˜ ao Paulo, Rua Doutor Carvalho de Mendonça, 144, 11070 - 100, Santos, SP, Brazil</mods:affiliation>
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<mods:namePart id="839368BAFC34A1BAC8D203D8A5129D3F">Nunes, Viviane Abreu</mods:namePart>
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<mods:affiliation id="EA052C858B940B3BB64D2863C69741E1">* & Department of Biotechnology, Universidade de S ˜ ao Paulo, Avenida Arlindo B´ettio, 1000, 03828 - 000, S ˜ ao Paulo, SP, Brazil</mods:affiliation>
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<mods:namePart id="C53865DEDB127323ECB4969F785D6A6D">Tanaka, Aparecida Sadae</mods:namePart>
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<mods:affiliation id="CEDBDF708A95882C3523B8069258B69C">Department of Biochemistry, Universidade Federal de S ˜ ao Paulo, Rua Trˆes de Maio, 100, 04044 - 020, S ˜ ao Paulo, SP, Brazil</mods:affiliation>
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<mods:namePart id="E038D1BB51C0DD928A5EA922AB32F74C">Araujo, Mariana da Silva</mods:namePart>
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<mods:affiliation id="69A2A0297D93C4ADD2C415F9DDC0B0EF">Department of Biochemistry, Universidade Federal de S ˜ ao Paulo, Rua Trˆes de Maio, 100, 04044 - 020, S ˜ ao Paulo, SP, Brazil</mods:affiliation>
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<mods:title id="3F40FE6AAC8CF98B2398181A9D483230">Phytochemistry</mods:title>
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<mods:date id="52E26887423AF83ED25C77C1845B0293">2021</mods:date>
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<mods:title id="4A9F10934334E1049A23E792421D98E2">112595</mods:title>
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<mods:number id="954B5C17DF470FA8545129DFB7C9C14A">2021-02-28</mods:number>
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<mods:number id="8995E176BE036ACC274FFDF3E9DB18B1">182</mods:number>
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<mods:classification id="3134F27E64EB346FCB4E4D65D1CFEF22">journal article</mods:classification>
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<mods:identifier id="239A7E2AA7F6D050D017CE9D1393D077" type="DOI">10.1016/j.phytochem.2020.112595</mods:identifier>
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<mods:identifier id="52DD5CD62077D920ADF0E3E529F93AF5" type="ISSN">1873-3700</mods:identifier>
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<treatment id="03FC87C47220FF89FB03107AFA78FD20" LSID="urn:lsid:plazi:treatment:03FC87C47220FF89FB03107AFA78FD20" httpUri="http://treatment.plazi.org/id/03FC87C47220FF89FB03107AFA78FD20" lastPageNumber="6" pageId="5" pageNumber="6">
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<subSubSection id="C34F65597220FF89FB03107AFE01F9CF" box="[100,425,1627,1646]" pageId="5" pageNumber="6" type="nomenclature">
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<paragraph id="8BEA36D27220FF89FB03107AFE01F9CF" blockId="5.[100,425,1627,1646]" box="[100,425,1627,1646]" pageId="5" pageNumber="6">
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<heading id="D0A281BE7220FF89FB03107AFE01F9CF" bold="true" box="[100,425,1627,1646]" fontSize="36" level="1" pageId="5" pageNumber="6" reason="1">
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<emphasis id="B921EAC07220FF89FB03107AFE01F9CF" bold="true" box="[100,425,1627,1646]" italics="true" pageId="5" pageNumber="6">
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4.5. Isolation of
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<taxonomicName id="4C554D517220FF89FA66107AFECEF9CF" ID-CoL="694XL" authority="Lam." box="[257,358,1627,1646]" class="Magnoliopsida" family="Fabaceae" genus="Caesalpinia" kingdom="Plantae" order="Fabales" pageId="5" pageNumber="6" phylum="Tracheophyta" rank="species" species="echinata">C. echinata</taxonomicName>
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mRNA
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</emphasis>
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<subSubSection id="C34F65597220FF89FBE310B2FA78FD20" pageId="5" pageNumber="6" type="description">
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<paragraph id="8BEA36D27220FF89FBE310B2FF19F893" blockId="5.[100,771,1683,1842]" pageId="5" pageNumber="6">
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The RNeasy Plant Mini kit was used to isolate the mRNA. Briefly,
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<taxonomicName id="4C554D517220FF89FB03108EFF6FF963" box="[100,199,1711,1730]" class="Magnoliopsida" family="Fabaceae" genus="Caesalpinia" kingdom="Plantae" order="Fabales" pageId="5" pageNumber="6" phylum="Tracheophyta" rank="species" species="echinata">
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<emphasis id="B921EAC07220FF89FB03108EFF6FF963" bold="true" box="[100,199,1711,1730]" italics="true" pageId="5" pageNumber="6">C. echinata</emphasis>
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seeds, harvest tree or eight weeks after flowering were frozen in liquid nitrogen.
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<quantity id="4CAD9B377220FF89FA7E10EAFECAF97E" box="[281,354,1739,1759]" metricMagnitude="-4" metricUnit="kg" metricValue="1.0" pageId="5" pageNumber="6" unit="mg" value="100.0">100 mg</quantity>
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of the resulting powder was incubated, according to the manufacturer, in a buffer containing guanidinium thiocyanate and β- mercaptoethanol, and then purified into a QIAshredder column.
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</paragraph>
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<emphasis id="B921EAC07220FF89FB031176FF43F827" bold="true" italics="true" pageId="5" pageNumber="6">
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4.6.
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<taxonomicName id="4C554D517220FF89FBF21176FF53F8C8" ID-CoL="694XL" authority="Lam." box="[149,251,1878,1898]" class="Magnoliopsida" family="Fabaceae" genus="Caesalpinia" kingdom="Plantae" order="Fabales" pageId="5" pageNumber="6" phylum="Tracheophyta" rank="species" species="echinata">C. echinata</taxonomicName>
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cDNA synthesis and amplification by polymerase chain reaction (PCR)
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</emphasis>
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</paragraph>
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<paragraph id="8BEA36D27220FF89FBE3118AFA78FD20" blockId="5.[132,770,1963,1982]" lastBlockId="5.[818,1488,145,641]" pageId="5" pageNumber="6">
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The cDNA synthesis was performed with the ImProm-IITM Reverse Transcriptase, total mRNA and the oligo primer race adapter 3
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as primer, as recommended by the manufacturer. The degenerate oligonucleotide ODN-CeEI (5
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TTY GTN GTN GAY ACN GAR GRN AAY YTN HTN CAR AAY GGN GG 3
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), based on the N-terminal amino acid sequence of CeEI (
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<bibRefCitation id="EFC44B237220FF89F8861722FB1CFEB7" author="Cruz-Silva, I. & Neuhof, C. & Gozzo, A. J. & Nunes, V. A. & Hirata, I. Y. & Sampaio, M. U. & Figueiredo-Ribeiro, R. de & Neuhof, H. & Araujo, M. S." box="[993,1204,259,279]" pageId="5" pageNumber="6" pagination="235 - 243" refId="ref9583" refString="Cruz-Silva, I., Neuhof, C., Gozzo, A. J., Nunes, V. A., Hirata, I. Y., Sampaio, M. U., Figueiredo-Ribeiro, R. de C., Neuhof, H., Araujo, M. S., 2013. Using a Caesalpinia echinata Lam. protease inhibitor as a tool for studying the roles of neutrophil elastase, cathepsin G and proteinase 3 in pulmonary edema. Phytochemistry 96, 235 - 243. https: // doi. org / 10.1016 / j. phytochem. 2013.09.025." type="journal article" year="2013">Cruz-Silva et al., 2013</bibRefCitation>
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), was used as forward primer and the Abridged Universal Amplification Primer - AUAP - (5
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<geneSequence id="5E48AE4A7220FF89FE11173EFBBAFEEF" pageId="5" pageNumber="6">GGC CAC GCG TCG ACT AGT AC</geneSequence>
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3
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) was used as reverse primer. Briefly, cDNA (2.0 μg) were incubated with Taq polymerase (5 U), oligonucleotide AUAP (50 pM), degenerate oligonucleotide (50 pM), dNTP mix (0.20 mM) e MgCl
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(2.5 mM), in a total volume of 50 μL. Reactions with only one oligonucleotide were also performed as control. It was used pre-denaturation (94
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C for 5 min), PCR amplification of 35 cycles (denaturing at 94
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C for 40 s, annealing at 68
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C for 40 s, and extension at 72
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C for 100 s), and final extension (72
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C for 5 min). The amplified DNA fragments were analyzed by gel agarose (1%) electrophoresis containing 1.27 μM ethidium bromide. The DNA fragment of 700 bp, corresponding to the CeEI DNA fragment, was purified in agarose using QIAEX II gel extraction kit and ligated into pGEM-T easy cloning vector.
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</paragraph>
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</subSubSection>
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</treatment>
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