Molecular Cloning and Expression Analysis of a Catalase Gene (NnCAT) from Nelumbo nucifera
Rapid amplification cDNA end (RACE) assay was established to achieve the complete cDNA sequence of a catalase gene (NnCAT) from Nelumbo nucifera. The obtained full-length cDNA was 1666 bp in size and contained a 1476-bp open reading frame. The 3D structural model of NnCAT was constructed by homology...
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| Veröffentlicht in: | Applied biochemistry and biotechnology 2015-11, Vol.177 (6), p.1216-1228 |
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| creator | Dong, Chen Zheng, Xingfei Diao, Ying Wang, Youwei Zhou, Mingquan Hu, Zhongli |
| description | Rapid amplification cDNA end (RACE) assay was established to achieve the complete cDNA sequence of a catalase gene (NnCAT) from Nelumbo nucifera. The obtained full-length cDNA was 1666 bp in size and contained a 1476-bp open reading frame. The 3D structural model of NnCAT was constructed by homology modeling. The putative NnCAT possessed all the main characteristic amino acid residues and motifs of catalase (CAT) protein family, and the phylogenetic analysis revealed that NnCAT grouped together with high plants. Moreover, recombinant NnCAT showed the CAT activity (758 U/mg) at room temperature, holding high activity during temperature range of 20–50 °C, then the optimal pH of recombinant protein was assessed from pH 4 to pH 11. Additionally, real-time PCR assay demonstrated that NnCAT mRNA was expressed in various tissues of N. nucifera, with the highest expression in young leaf and lowest level in the root, and mRNA level of NnCAT was significantly augmented in response to short-time mechanical wounding. Different expression pattern of NnCAT gene suggested that NnCAT probably played a defensive role in the initial stages of oxidative stress, regulating the level of reactive oxygen species (ROS) by extracellular stimuli such as short-time mechanical wounding. |
| doi_str_mv | 10.1007/s12010-015-1808-7 |
| format | Article |
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The obtained full-length cDNA was 1666 bp in size and contained a 1476-bp open reading frame. The 3D structural model of NnCAT was constructed by homology modeling. The putative NnCAT possessed all the main characteristic amino acid residues and motifs of catalase (CAT) protein family, and the phylogenetic analysis revealed that NnCAT grouped together with high plants. Moreover, recombinant NnCAT showed the CAT activity (758 U/mg) at room temperature, holding high activity during temperature range of 20–50 °C, then the optimal pH of recombinant protein was assessed from pH 4 to pH 11. Additionally, real-time PCR assay demonstrated that NnCAT mRNA was expressed in various tissues of N. nucifera, with the highest expression in young leaf and lowest level in the root, and mRNA level of NnCAT was significantly augmented in response to short-time mechanical wounding. Different expression pattern of NnCAT gene suggested that NnCAT probably played a defensive role in the initial stages of oxidative stress, regulating the level of reactive oxygen species (ROS) by extracellular stimuli such as short-time mechanical wounding.</description><identifier>ISSN: 0273-2289</identifier><identifier>EISSN: 1559-0291</identifier><identifier>DOI: 10.1007/s12010-015-1808-7</identifier><identifier>PMID: 26299377</identifier><language>eng</language><publisher>New York: Springer US</publisher><subject>ambient temperature ; amino acid composition ; Amino acids ; Aquatic plants ; Biochemistry ; Biotechnology ; catalase ; Catalase - biosynthesis ; Catalase - chemistry ; Catalase - genetics ; Chemistry ; Chemistry and Materials Science ; Cloning ; Cloning, Molecular ; complementary DNA ; Enzymes ; Gene expression ; Gene Expression Regulation, Enzymologic - physiology ; Gene Expression Regulation, Plant - physiology ; genes ; leaves ; messenger RNA ; Models, Molecular ; molecular cloning ; Nelumbo - enzymology ; Nelumbo - genetics ; Nelumbo nucifera ; open reading frames ; Oxidative stress ; phylogeny ; Plant Proteins - biosynthesis ; Plant Proteins - chemistry ; Plant Proteins - genetics ; Protein Structure, Tertiary ; quantitative polymerase chain reaction ; rapid amplification of cDNA ends ; reactive oxygen species ; recombinant proteins ; tissues</subject><ispartof>Applied biochemistry and biotechnology, 2015-11, Vol.177 (6), p.1216-1228</ispartof><rights>Springer Science+Business Media New York 2015</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c536t-38a2a082af13ad80c3a426eb9fc54c08a258513f84f42d4f647ef4c78b14ef113</citedby><cites>FETCH-LOGICAL-c536t-38a2a082af13ad80c3a426eb9fc54c08a258513f84f42d4f647ef4c78b14ef113</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s12010-015-1808-7$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s12010-015-1808-7$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26299377$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Dong, Chen</creatorcontrib><creatorcontrib>Zheng, Xingfei</creatorcontrib><creatorcontrib>Diao, Ying</creatorcontrib><creatorcontrib>Wang, Youwei</creatorcontrib><creatorcontrib>Zhou, Mingquan</creatorcontrib><creatorcontrib>Hu, Zhongli</creatorcontrib><title>Molecular Cloning and Expression Analysis of a Catalase Gene (NnCAT) from Nelumbo nucifera</title><title>Applied biochemistry and biotechnology</title><addtitle>Appl Biochem Biotechnol</addtitle><addtitle>Appl Biochem Biotechnol</addtitle><description>Rapid amplification cDNA end (RACE) assay was established to achieve the complete cDNA sequence of a catalase gene (NnCAT) from Nelumbo nucifera. The obtained full-length cDNA was 1666 bp in size and contained a 1476-bp open reading frame. The 3D structural model of NnCAT was constructed by homology modeling. The putative NnCAT possessed all the main characteristic amino acid residues and motifs of catalase (CAT) protein family, and the phylogenetic analysis revealed that NnCAT grouped together with high plants. Moreover, recombinant NnCAT showed the CAT activity (758 U/mg) at room temperature, holding high activity during temperature range of 20–50 °C, then the optimal pH of recombinant protein was assessed from pH 4 to pH 11. Additionally, real-time PCR assay demonstrated that NnCAT mRNA was expressed in various tissues of N. nucifera, with the highest expression in young leaf and lowest level in the root, and mRNA level of NnCAT was significantly augmented in response to short-time mechanical wounding. Different expression pattern of NnCAT gene suggested that NnCAT probably played a defensive role in the initial stages of oxidative stress, regulating the level of reactive oxygen species (ROS) by extracellular stimuli such as short-time mechanical wounding.</description><subject>ambient temperature</subject><subject>amino acid composition</subject><subject>Amino acids</subject><subject>Aquatic plants</subject><subject>Biochemistry</subject><subject>Biotechnology</subject><subject>catalase</subject><subject>Catalase - biosynthesis</subject><subject>Catalase - chemistry</subject><subject>Catalase - genetics</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Cloning</subject><subject>Cloning, Molecular</subject><subject>complementary DNA</subject><subject>Enzymes</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Enzymologic - physiology</subject><subject>Gene Expression Regulation, Plant - physiology</subject><subject>genes</subject><subject>leaves</subject><subject>messenger RNA</subject><subject>Models, Molecular</subject><subject>molecular cloning</subject><subject>Nelumbo - enzymology</subject><subject>Nelumbo - genetics</subject><subject>Nelumbo nucifera</subject><subject>open reading frames</subject><subject>Oxidative stress</subject><subject>phylogeny</subject><subject>Plant Proteins - biosynthesis</subject><subject>Plant Proteins - chemistry</subject><subject>Plant Proteins - genetics</subject><subject>Protein Structure, Tertiary</subject><subject>quantitative polymerase chain reaction</subject><subject>rapid amplification of cDNA ends</subject><subject>reactive oxygen species</subject><subject>recombinant proteins</subject><subject>tissues</subject><issn>0273-2289</issn><issn>1559-0291</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNkT1vFDEQhi1ERC6BH0ADlmhCscFjr9d2eVqFBCkfBUlDY_l89mkjr33YtxL593i1ASEKRDXFPO870jwIvQVyDoSITwUoAdIQ4A1IIhvxAq2Ac9UQquAlWhEqWEOpVMfopJRHQoBKLl6hY9pRpZgQK_TtJgVnp2Ay7kOKQ9xhE7f44sc-u1KGFPE6mvBUhoKTxwb35mCCKQ5fuujw2W3s1_cfsc9pxLcuTOMm4TjZwbtsXqMjb0Jxb57nKXr4fHHfXzXXd5df-vV1YznrDg2ThhoiqfHAzFYSy0xLO7dR3vLWkrrlkgPzsvUt3ba-a4XzrRVyA63zAOwUnS29-5y-T64c9DgU60Iw0aWpaBCMK0o5Zf-DglD1bXPrh7_QxzTl-ouZokqwDqSoFCyUzamU7Lze52E0-UkD0bMjvTjS1ZGeHek58-65edqMbvs78UtKBegClLqKO5f_OP2P1vdLyJukzS4PRT98rVBHqnaQHbCfQo-ieA</recordid><startdate>20151101</startdate><enddate>20151101</enddate><creator>Dong, Chen</creator><creator>Zheng, Xingfei</creator><creator>Diao, Ying</creator><creator>Wang, Youwei</creator><creator>Zhou, Mingquan</creator><creator>Hu, Zhongli</creator><general>Springer US</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>SOI</scope><scope>7X8</scope><scope>7QO</scope></search><sort><creationdate>20151101</creationdate><title>Molecular Cloning and Expression Analysis of a Catalase Gene (NnCAT) from Nelumbo nucifera</title><author>Dong, Chen ; Zheng, Xingfei ; Diao, Ying ; Wang, Youwei ; Zhou, Mingquan ; Hu, Zhongli</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c536t-38a2a082af13ad80c3a426eb9fc54c08a258513f84f42d4f647ef4c78b14ef113</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>ambient temperature</topic><topic>amino acid composition</topic><topic>Amino acids</topic><topic>Aquatic plants</topic><topic>Biochemistry</topic><topic>Biotechnology</topic><topic>catalase</topic><topic>Catalase - biosynthesis</topic><topic>Catalase - chemistry</topic><topic>Catalase - genetics</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Cloning</topic><topic>Cloning, Molecular</topic><topic>complementary DNA</topic><topic>Enzymes</topic><topic>Gene expression</topic><topic>Gene Expression Regulation, Enzymologic - physiology</topic><topic>Gene Expression Regulation, Plant - physiology</topic><topic>genes</topic><topic>leaves</topic><topic>messenger RNA</topic><topic>Models, Molecular</topic><topic>molecular cloning</topic><topic>Nelumbo - enzymology</topic><topic>Nelumbo - genetics</topic><topic>Nelumbo nucifera</topic><topic>open reading frames</topic><topic>Oxidative stress</topic><topic>phylogeny</topic><topic>Plant Proteins - biosynthesis</topic><topic>Plant Proteins - chemistry</topic><topic>Plant Proteins - genetics</topic><topic>Protein Structure, Tertiary</topic><topic>quantitative polymerase chain reaction</topic><topic>rapid amplification of cDNA ends</topic><topic>reactive oxygen species</topic><topic>recombinant proteins</topic><topic>tissues</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Dong, Chen</creatorcontrib><creatorcontrib>Zheng, Xingfei</creatorcontrib><creatorcontrib>Diao, Ying</creatorcontrib><creatorcontrib>Wang, Youwei</creatorcontrib><creatorcontrib>Zhou, Mingquan</creatorcontrib><creatorcontrib>Hu, Zhongli</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Environment Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>Environment Abstracts</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><jtitle>Applied biochemistry and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Dong, Chen</au><au>Zheng, Xingfei</au><au>Diao, Ying</au><au>Wang, Youwei</au><au>Zhou, Mingquan</au><au>Hu, Zhongli</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular Cloning and Expression Analysis of a Catalase Gene (NnCAT) from Nelumbo nucifera</atitle><jtitle>Applied biochemistry and biotechnology</jtitle><stitle>Appl Biochem Biotechnol</stitle><addtitle>Appl Biochem Biotechnol</addtitle><date>2015-11-01</date><risdate>2015</risdate><volume>177</volume><issue>6</issue><spage>1216</spage><epage>1228</epage><pages>1216-1228</pages><issn>0273-2289</issn><eissn>1559-0291</eissn><abstract>Rapid amplification cDNA end (RACE) assay was established to achieve the complete cDNA sequence of a catalase gene (NnCAT) from Nelumbo nucifera. The obtained full-length cDNA was 1666 bp in size and contained a 1476-bp open reading frame. The 3D structural model of NnCAT was constructed by homology modeling. The putative NnCAT possessed all the main characteristic amino acid residues and motifs of catalase (CAT) protein family, and the phylogenetic analysis revealed that NnCAT grouped together with high plants. Moreover, recombinant NnCAT showed the CAT activity (758 U/mg) at room temperature, holding high activity during temperature range of 20–50 °C, then the optimal pH of recombinant protein was assessed from pH 4 to pH 11. Additionally, real-time PCR assay demonstrated that NnCAT mRNA was expressed in various tissues of N. nucifera, with the highest expression in young leaf and lowest level in the root, and mRNA level of NnCAT was significantly augmented in response to short-time mechanical wounding. Different expression pattern of NnCAT gene suggested that NnCAT probably played a defensive role in the initial stages of oxidative stress, regulating the level of reactive oxygen species (ROS) by extracellular stimuli such as short-time mechanical wounding.</abstract><cop>New York</cop><pub>Springer US</pub><pmid>26299377</pmid><doi>10.1007/s12010-015-1808-7</doi><tpages>13</tpages></addata></record> |
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| subjects | ambient temperature amino acid composition Amino acids Aquatic plants Biochemistry Biotechnology catalase Catalase - biosynthesis Catalase - chemistry Catalase - genetics Chemistry Chemistry and Materials Science Cloning Cloning, Molecular complementary DNA Enzymes Gene expression Gene Expression Regulation, Enzymologic - physiology Gene Expression Regulation, Plant - physiology genes leaves messenger RNA Models, Molecular molecular cloning Nelumbo - enzymology Nelumbo - genetics Nelumbo nucifera open reading frames Oxidative stress phylogeny Plant Proteins - biosynthesis Plant Proteins - chemistry Plant Proteins - genetics Protein Structure, Tertiary quantitative polymerase chain reaction rapid amplification of cDNA ends reactive oxygen species recombinant proteins tissues |
| title | Molecular Cloning and Expression Analysis of a Catalase Gene (NnCAT) from Nelumbo nucifera |
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