Reference genes identified in the silkworm Bombyx mori during metamorphism based on oligonucleotide microarray and confirmed by qRT‐PCR
Identifieur interne : 000603 ( Istex/Corpus ); précédent : 000602; suivant : 000604Reference genes identified in the silkworm Bombyx mori during metamorphism based on oligonucleotide microarray and confirmed by qRT‐PCR
Auteurs : Gen-Hong Wang ; Qing-You Xia ; Dao-Jun Cheng ; Jun Duan ; Ping Zhao ; Jie Chen ; Li ZhuSource :
- Insect Science [ 1672-9609 ] ; 2008-10.
English descriptors
- Teeft :
- Actin, Authors journal compilation institute, Bombyx, Bombyx mori, Candidate reference genes, Chinese academy, Control genes, Cytoplasmic, Cytoplasmic actin, Data normalization, Developmental stage, Developmental stages, Expression levels, Expression pattern, Expression stability, Fifth instar, Further validation, Gene, Gene expression normalization, Gene expression quantification, Genome, Genorm, Genorm software, Housekeeping genes, Insect science, Instar, Internal controls, Microarray, Mori, Mrna, Normalization, Normalization factor, Normfinder, Oligonucleotide microarray, Optimal number, Primer, Program normfinder, Proteasome beta subunit, Protein synthesis, Reference gene, Reference gene candidates, Reference genes, Ribosomal, Ribosomal protein, Ribosomal protein genes, Silkworm, Silkworm bombyx mori, Stable genes, Stable reference genes, Subunit, Translation initiation factor, Translation initiation factors, Wang, Zhang, Zoology.
Abstract
Gene expression quantification at mRNA level is very important for post‐genomic studies, as gene expression level is the reflection of the special biological function of the target gene. Methods used for gene expression quantification, such as microarray or quantitative real‐time polymerase chain reaction (qRT‐PCR), require stable expressed reference genes. Thus, finding suitable control genes is essential for gene quantification. In this study, a genome‐wide survey of reference genes during metamorphism was performed on silkworm Bombyx mori. Twelve genes were chosen as putative reference genes based on a whole genome oligonucleotide microarray normalized by external controls. Then, qRT‐PCR was employed for further validation and selection of potential reference gene candidates. The results were analyzed, and stable genes were selected using geNorm 3.4 and NormFinder software. Finally, considering factors from every aspect, translation initiation factor 4A, translation initiation factor 3 subunit 4, and translation initiation factor 3 subunit 5 (represented by sw22934, swl4876, and swl3956) were selected as reliable internal controls across the examined developmental stages, while cytoplasmic actin (sw22671), the commonly used reference gene in a previous study was shown to vary drastically throughout the examined developmental stages. For future research, we recommend the use of the geometric mean of those three stable reference genes as an accurate normalization factor for data normalization of different developmental stages during metamorphism.
Url:
DOI: 10.1111/j.1744-7917.2008.00227.x
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ISTEX:E2A351E4100C5023D8E964D986BEA645E7B0DBCALe document en format XML
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China</mods:affiliation></affiliation><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Cheng, Dao Un" sort="Cheng, Dao Un" uniqKey="Cheng D" first="Dao-Jun" last="Cheng">Dao-Jun Cheng</name><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Duan, Jun" sort="Duan, Jun" uniqKey="Duan J" first="Jun" last="Duan">Jun Duan</name><affiliation><mods:affiliation>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</mods:affiliation></affiliation><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Zhao, Ping" sort="Zhao, Ping" uniqKey="Zhao P" first="Ping" last="Zhao">Ping Zhao</name><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Chen, Jie" sort="Chen, Jie" uniqKey="Chen J" first="Jie" last="Chen">Jie Chen</name><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Zhu, Li" sort="Zhu, Li" uniqKey="Zhu L" first="Li" last="Zhu">Li Zhu</name><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno 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Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Xia, Qing Ou" sort="Xia, Qing Ou" uniqKey="Xia Q" first="Qing-You" last="Xia">Qing-You Xia</name><affiliation><mods:affiliation>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</mods:affiliation></affiliation><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Cheng, Dao Un" sort="Cheng, Dao Un" uniqKey="Cheng D" first="Dao-Jun" last="Cheng">Dao-Jun Cheng</name><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Duan, Jun" sort="Duan, Jun" uniqKey="Duan J" first="Jun" last="Duan">Jun Duan</name><affiliation><mods:affiliation>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</mods:affiliation></affiliation><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Zhao, Ping" sort="Zhao, Ping" uniqKey="Zhao P" first="Ping" last="Zhao">Ping Zhao</name><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Chen, Jie" sort="Chen, Jie" uniqKey="Chen J" first="Jie" last="Chen">Jie Chen</name><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author><author><name sortKey="Zhu, Li" sort="Zhu, Li" uniqKey="Zhu L" first="Li" last="Zhu">Li Zhu</name><affiliation><mods:affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Insect Science</title><title level="j" type="alt">INSECT SCIENCE</title><idno type="ISSN">1672-9609</idno><idno type="eISSN">1744-7917</idno><imprint><biblScope unit="vol">15</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="405">405</biblScope><biblScope unit="page" to="413">413</biblScope><biblScope unit="page-count">9</biblScope><publisher>Blackwell Publishing Ltd</publisher><pubPlace>Oxford, UK</pubPlace><date type="published" when="2008-10">2008-10</date></imprint><idno type="ISSN">1672-9609</idno></series></biblStruct></sourceDesc><seriesStmt><idno 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factor</term><term>Normfinder</term><term>Oligonucleotide microarray</term><term>Optimal number</term><term>Primer</term><term>Program normfinder</term><term>Proteasome beta subunit</term><term>Protein synthesis</term><term>Reference gene</term><term>Reference gene candidates</term><term>Reference genes</term><term>Ribosomal</term><term>Ribosomal protein</term><term>Ribosomal protein genes</term><term>Silkworm</term><term>Silkworm bombyx mori</term><term>Stable genes</term><term>Stable reference genes</term><term>Subunit</term><term>Translation initiation factor</term><term>Translation initiation factors</term><term>Wang</term><term>Zhang</term><term>Zoology</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Gene expression quantification at mRNA level is very important for post‐genomic studies, as gene expression level is the reflection of the special biological function of the target gene. Methods used for gene expression quantification, such as microarray or quantitative real‐time polymerase chain reaction (qRT‐PCR), require stable expressed reference genes. Thus, finding suitable control genes is essential for gene quantification. In this study, a genome‐wide survey of reference genes during metamorphism was performed on silkworm Bombyx mori. Twelve genes were chosen as putative reference genes based on a whole genome oligonucleotide microarray normalized by external controls. Then, qRT‐PCR was employed for further validation and selection of potential reference gene candidates. The results were analyzed, and stable genes were selected using geNorm 3.4 and NormFinder software. Finally, considering factors from every aspect, translation initiation factor 4A, translation initiation factor 3 subunit 4, and translation initiation factor 3 subunit 5 (represented by sw22934, swl4876, and swl3956) were selected as reliable internal controls across the examined developmental stages, while cytoplasmic actin (sw22671), the commonly used reference gene in a previous study was shown to vary drastically throughout the examined developmental stages. For future research, we recommend the use of the geometric mean of those three stable reference genes as an accurate normalization factor for data normalization of different developmental stages during metamorphism.</div></front></TEI><istex><corpusName>wiley</corpusName><keywords><teeft><json:string>normalization</json:string><json:string>reference genes</json:string><json:string>mori</json:string><json:string>silkworm</json:string><json:string>actin</json:string><json:string>microarray</json:string><json:string>cytoplasmic</json:string><json:string>genorm</json:string><json:string>cytoplasmic actin</json:string><json:string>wang</json:string><json:string>bombyx</json:string><json:string>normfinder</json:string><json:string>subunit</json:string><json:string>ribosomal</json:string><json:string>translation initiation factor</json:string><json:string>primer</json:string><json:string>insect science</json:string><json:string>chinese academy</json:string><json:string>authors journal 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China</json:string></affiliations></json:item><json:item><name>Qing‐You Xia</name><affiliations><json:string>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</json:string><json:string>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</json:string></affiliations></json:item><json:item><name>Dao‐Jun Cheng</name><affiliations><json:string>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</json:string></affiliations></json:item><json:item><name>Jun Duan</name><affiliations><json:string>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</json:string><json:string>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</json:string></affiliations></json:item><json:item><name>Ping Zhao</name><affiliations><json:string>The 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gene</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>qRT‐PCR</value></json:item></subject><articleId><json:string>INS227</json:string></articleId><arkIstex>ark:/67375/WNG-F0LXHWKL-F</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>article</json:string></originalGenre><abstract>Gene expression quantification at mRNA level is very important for post‐genomic studies, as gene expression level is the reflection of the special biological function of the target gene. Methods used for gene expression quantification, such as microarray or quantitative real‐time polymerase chain reaction (qRT‐PCR), require stable expressed reference genes. Thus, finding suitable control genes is essential for gene quantification. In this study, a genome‐wide survey of reference genes during metamorphism was performed on silkworm Bombyx mori. Twelve genes were chosen as putative reference genes based on a whole genome oligonucleotide microarray normalized by external controls. Then, qRT‐PCR was employed for further validation and selection of potential reference gene candidates. The results were analyzed, and stable genes were selected using geNorm 3.4 and NormFinder software. Finally, considering factors from every aspect, translation initiation factor 4A, translation initiation factor 3 subunit 4, and translation initiation factor 3 subunit 5 (represented by sw22934, swl4876, and swl3956) were selected as reliable internal controls across the examined developmental stages, while cytoplasmic actin (sw22671), the commonly used reference gene in a previous study was shown to vary drastically throughout the examined developmental stages. For future research, we recommend the use of the geometric mean of those three stable reference genes as an accurate normalization factor for data normalization of different developmental stages during metamorphism.</abstract><qualityIndicators><score>9.184</score><pdfWordCount>4568</pdfWordCount><pdfCharCount>30478</pdfCharCount><pdfVersion>1.4</pdfVersion><pdfPageCount>9</pdfPageCount><pdfPageSize>595.3 x 779.55 pts</pdfPageSize><pdfWordsPerPage>508</pdfWordsPerPage><pdfText>true</pdfText><refBibsNative>true</refBibsNative><abstractWordCount>218</abstractWordCount><abstractCharCount>1573</abstractCharCount><keywordCount>5</keywordCount></qualityIndicators><title>Reference genes identified in the silkworm Bombyx mori during metamorphism based on oligonucleotide microarray and confirmed by qRT‐PCR</title><genre><json:string>article</json:string></genre><host><title>Insect Science</title><language><json:string>unknown</json:string></language><doi><json:string>10.1111/(ISSN)1744-7917</json:string></doi><issn><json:string>1672-9609</json:string></issn><eissn><json:string>1744-7917</json:string></eissn><publisherId><json:string>INS</json:string></publisherId><volume>15</volume><issue>5</issue><pages><first>405</first><last>413</last><total>9</total></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>2008</json:string></date><geogName></geogName><orgName><json:string>Institute of Zoology, Chinese Academy of Sciences Insect Science</json:string><json:string>Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing</json:string><json:string>Chongqing University</json:string><json:string>Institute of Agriculture and Life Sciences</json:string><json:string>Institute of Zoology, Chinese Academy of Sciences, Insect Science</json:string><json:string>Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China Abstract Gene</json:string><json:string>Tech Research and Development Program of China</json:string></orgName><orgName_funder></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Li Zhu</json:string><json:string>Nano Drop</json:string><json:string>Foster Table</json:string><json:string>Jie Chen</json:string><json:string>Dao-Jun Cheng</json:string><json:string>Qing-You Xia</json:string><json:string>Jun Duan</json:string></persName><placeName><json:string>Germany</json:string><json:string>Beijing</json:string><json:string>China</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>Xia et al.</json:string><json:string>Dhawan & Gopinathan, 2002</json:string><json:string>Mange et al., 1996</json:string><json:string>Eisenberg & Levanon, 2003</json:string><json:string>Mamo et al., 2007</json:string><json:string>Mackay et al., 2002</json:string><json:string>Livak & Schmittgen, 2001</json:string><json:string>Xia et al., 2006</json:string><json:string>de Kok et al., 2005</json:string><json:string>Wang et al., 2005</json:string><json:string>Thellin et al., 1999</json:string><json:string>Radonic et al., 2005</json:string><json:string>Vandesomple et al. 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(2003)</json:string><json:string>Labuhn & Brack, 1997</json:string><json:string>Shikama & Brack, 1996</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/WNG-F0LXHWKL-F</json:string></ark><categories><wos><json:string>1 - science</json:string><json:string>2 - entomology</json:string></wos><scienceMetrix><json:string>1 - natural sciences</json:string><json:string>2 - biology</json:string><json:string>3 - entomology</json:string></scienceMetrix><scopus><json:string>1 - Life Sciences</json:string><json:string>2 - Agricultural and Biological Sciences</json:string><json:string>3 - Insect Science</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Biochemistry, Genetics and Molecular Biology</json:string><json:string>3 - General Biochemistry, Genetics and Molecular Biology</json:string><json:string>1 - Life Sciences</json:string><json:string>2 - Agricultural and Biological Sciences</json:string><json:string>3 - Agronomy and Crop 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type="institution">Chongqing University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><affiliation><orgName type="laboratory">The Key Sericultural Laboratory of Agricultural Ministry</orgName><orgName type="department">School of Biotechnology</orgName><orgName type="institution">Southwest University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation></author><author xml:id="author-0001"><persName><forename type="first">Qing‐You</forename><surname>Xia</surname></persName><affiliation><orgName type="institution">Institute of Agriculture and Life Sciences</orgName><orgName type="institution">Chongqing University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><affiliation><orgName type="laboratory">The Key Sericultural Laboratory of Agricultural Ministry</orgName><orgName type="department">School of Biotechnology</orgName><orgName type="institution">Southwest University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation></author><author xml:id="author-0002"><persName><forename type="first">Dao‐Jun</forename><surname>Cheng</surname></persName><affiliation><orgName type="laboratory">The Key Sericultural Laboratory of Agricultural Ministry</orgName><orgName type="department">School of Biotechnology</orgName><orgName type="institution">Southwest University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation></author><author xml:id="author-0003"><persName><forename type="first">Jun</forename><surname>Duan</surname></persName><affiliation><orgName type="institution">Institute of Agriculture and Life Sciences</orgName><orgName type="institution">Chongqing University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation><affiliation><orgName type="laboratory">The Key Sericultural Laboratory of Agricultural Ministry</orgName><orgName type="department">School of Biotechnology</orgName><orgName type="institution">Southwest University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation></author><author xml:id="author-0004"><persName><forename type="first">Ping</forename><surname>Zhao</surname></persName><affiliation><orgName type="laboratory">The Key Sericultural Laboratory of Agricultural Ministry</orgName><orgName type="department">School of Biotechnology</orgName><orgName type="institution">Southwest University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation></author><author xml:id="author-0005"><persName><forename type="first">Jie</forename><surname>Chen</surname></persName><affiliation><orgName type="laboratory">The Key Sericultural Laboratory of Agricultural Ministry</orgName><orgName type="department">School of Biotechnology</orgName><orgName type="institution">Southwest University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation></author><author xml:id="author-0006"><persName><forename type="first">Li</forename><surname>Zhu</surname></persName><affiliation><orgName type="laboratory">The Key Sericultural Laboratory of Agricultural Ministry</orgName><orgName type="department">School of Biotechnology</orgName><orgName type="institution">Southwest University</orgName><address><addrLine>Chongqing</addrLine><addrLine>China</addrLine><country key="CN" xml:lang="en">CHINA</country></address></affiliation></author><idno type="istex">E2A351E4100C5023D8E964D986BEA645E7B0DBCA</idno><idno type="ark">ark:/67375/WNG-F0LXHWKL-F</idno><idno type="DOI">10.1111/j.1744-7917.2008.00227.x</idno><idno type="unit">INS227</idno><idno type="toTypesetVersion">file:INS.INS227.pdf</idno></analytic><monogr><title level="j" type="main">Insect Science</title><title level="j" type="alt">INSECT SCIENCE</title><idno type="pISSN">1672-9609</idno><idno type="eISSN">1744-7917</idno><idno type="book-DOI">10.1111/(ISSN)1744-7917</idno><idno type="book-part-DOI">10.1111/ins.2008.15.issue-5</idno><idno type="product">INS</idno><idno type="publisherDivision">ST</idno><imprint><biblScope unit="vol">15</biblScope><biblScope unit="issue">5</biblScope><biblScope unit="page" from="405">405</biblScope><biblScope unit="page" to="413">413</biblScope><biblScope unit="page-count">9</biblScope><publisher>Blackwell Publishing Ltd</publisher><pubPlace>Oxford, UK</pubPlace><date type="published" when="2008-10"></date></imprint></monogr></biblStruct></sourceDesc></fileDesc><encodingDesc><schemaRef type="ODD" url="https://xml-schema.delivery.istex.fr/tei-istex.odd"></schemaRef><appInfo><application ident="pub2tei" version="1.0.10" when="2019-12-20"><label>pub2TEI-ISTEX</label><desc>A set of style sheets for converting XML documents encoded in various scientific publisher formats into a common TEI format.
<ref target="http://www.tei-c.org/">We use TEI</ref></desc></application></appInfo></encodingDesc><profileDesc><abstract xml:lang="en" style="main"><head>Abstract</head><p>Gene expression quantification at mRNA level is very important for post‐genomic studies, as gene expression level is the reflection of the special biological function of the target gene. Methods used for gene expression quantification, such as microarray or quantitative real‐time polymerase chain reaction (qRT‐PCR), require stable expressed reference genes. Thus, finding suitable control genes is essential for gene quantification. In this study, a genome‐wide survey of reference genes during metamorphism was performed on silkworm <hi rend="italic">Bombyx mori.</hi> Twelve genes were chosen as putative reference genes based on a whole genome oligonucleotide microarray normalized by external controls. Then, qRT‐PCR was employed for further validation and selection of potential reference gene candidates. The results were analyzed, and stable genes were selected using geNorm 3.4 and NormFinder software. Finally, considering factors from every aspect, translation initiation factor 4A, translation initiation factor 3 subunit 4, and translation initiation factor 3 subunit 5 (represented by sw22934, swl4876, and swl3956) were selected as reliable internal controls across the examined developmental stages, while cytoplasmic actin (sw22671), the commonly used reference gene in a previous study was shown to vary drastically throughout the examined developmental stages. For future research, we recommend the use of the geometric mean of those three stable reference genes as an accurate normalization factor for data normalization of different developmental stages during metamorphism.</p></abstract><textClass><keywords xml:lang="en"><term xml:id="k1"><hi rend="italic">Bombyx mori</hi></term><term xml:id="k2">microarray</term><term xml:id="k3">nomalization factor</term><term xml:id="k4">reference gene</term><term xml:id="k5">qRT‐PCR</term></keywords><keywords rend="tocHeading1"><term>Original articles</term></keywords></textClass><langUsage><language ident="en"></language></langUsage></profileDesc><revisionDesc><change when="2019-12-20" who="#istex" xml:id="pub2tei">formatting</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/WNG-F0LXHWKL-F/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Wiley, elements deleted: body"><istex:xmlDeclaration>version="1.0" encoding="UTF-8" standalone="yes"</istex:xmlDeclaration><istex:document><component version="2.0" type="serialArticle" xml:lang="en"><header><publicationMeta level="product"><publisherInfo><publisherName>Blackwell Publishing Ltd</publisherName><publisherLoc>Oxford, UK</publisherLoc></publisherInfo><doi origin="wiley" registered="yes">10.1111/(ISSN)1744-7917</doi><issn type="print">1672-9609</issn><issn type="electronic">1744-7917</issn><idGroup><id type="product" value="INS"></id><id type="publisherDivision" value="ST"></id></idGroup><titleGroup><title type="main" sort="INSECT SCIENCE">Insect Science</title></titleGroup></publicationMeta><publicationMeta level="part" position="10005"><doi origin="wiley">10.1111/ins.2008.15.issue-5</doi><numberingGroup><numbering type="journalVolume" number="15">15</numbering><numbering type="journalIssue" number="5">5</numbering></numberingGroup><coverDate startDate="2008-10">October 2008</coverDate></publicationMeta><publicationMeta level="unit" type="article" position="3" status="forIssue"><doi origin="wiley">10.1111/j.1744-7917.2008.00227.x</doi><idGroup><id type="unit" value="INS227"></id></idGroup><countGroup><count type="pageTotal" number="9"></count></countGroup><titleGroup><title type="tocHeading1">Original articles</title></titleGroup><copyright>© 2008 The Authors Journal compilation © Institute of Zoology, Chinese Academy of Sciences</copyright><eventGroup><event type="firstOnline" date="2008-10-17"></event><event type="publishedOnlineFinalForm" date="2008-10-17"></event><event type="xmlConverted" agent="Converter:BPG_TO_WML3G version:2.3.2 mode:FullText source:HeaderRef result:HeaderRef" date="2010-03-07"></event><event type="xmlConverted" agent="Converter:WILEY_ML3G_TO_WILEY_ML3GV2 version:3.8.8" date="2014-01-29"></event><event type="xmlConverted" agent="Converter:WML3G_To_WML3G version:4.1.7 mode:FullText,remove_FC" date="2014-10-23"></event></eventGroup><numberingGroup><numbering type="pageFirst" number="405">405</numbering><numbering type="pageLast" number="413">413</numbering></numberingGroup><correspondenceTo> Qing‐You Xia, Institute of Agriculture and Life Sciences, Chongqing University, Chongqing 400030, and the Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing 400716, China. Tel: +86 23 68250099; fax: +86 23 68251128; email: <email>xiaqy@swu.edu.en</email></correspondenceTo><linkGroup><link type="toTypesetVersion" href="file:INS.INS227.pdf"></link></linkGroup></publicationMeta><contentMeta><unparsedEditorialHistory>Accepted May 7, 2008</unparsedEditorialHistory><countGroup><count type="referenceTotal" number="29"></count><count type="linksCrossRef" number="7"></count></countGroup><titleGroup><title type="main">Reference genes identified in the silkworm <i>Bombyx mori</i> during metamorphism based on oligonucleotide microarray and confirmed by qRT‐PCR</title></titleGroup><creators><creator creatorRole="author" xml:id="cr1" affiliationRef="#a1 #a2"><personName><givenNames>Gen‐Hong</givenNames><familyName>Wang</familyName></personName></creator><creator creatorRole="author" xml:id="cr2" affiliationRef="#a1 #a2"><personName><givenNames>Qing‐You</givenNames><familyName>Xia</familyName></personName></creator><creator creatorRole="author" xml:id="cr3" affiliationRef="#a2"><personName><givenNames>Dao‐Jun</givenNames><familyName>Cheng</familyName></personName></creator><creator creatorRole="author" xml:id="cr4" affiliationRef="#a1 #a2"><personName><givenNames>Jun</givenNames><familyName>Duan</familyName></personName></creator><creator creatorRole="author" xml:id="cr5" affiliationRef="#a2"><personName><givenNames>Ping</givenNames><familyName>Zhao</familyName></personName></creator><creator creatorRole="author" xml:id="cr6" affiliationRef="#a2"><personName><givenNames>Jie</givenNames><familyName>Chen</familyName></personName></creator><creator creatorRole="author" xml:id="cr7" affiliationRef="#a2"><personName><givenNames>Li</givenNames><familyName>Zhu</familyName></personName></creator></creators><affiliationGroup><affiliation xml:id="a1" countryCode="CN"><unparsedAffiliation>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</unparsedAffiliation></affiliation><affiliation xml:id="a2" countryCode="CN"><unparsedAffiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</unparsedAffiliation></affiliation></affiliationGroup><keywordGroup xml:lang="en"><keyword xml:id="k1"><i>Bombyx mori</i></keyword><keyword xml:id="k2">microarray</keyword><keyword xml:id="k3">nomalization factor</keyword><keyword xml:id="k4">reference gene</keyword><keyword xml:id="k5">qRT‐PCR</keyword></keywordGroup><abstractGroup><abstract type="main" xml:lang="en"><title type="main">Abstract</title><p>Gene expression quantification at mRNA level is very important for post‐genomic studies, as gene expression level is the reflection of the special biological function of the target gene. Methods used for gene expression quantification, such as microarray or quantitative real‐time polymerase chain reaction (qRT‐PCR), require stable expressed reference genes. Thus, finding suitable control genes is essential for gene quantification. In this study, a genome‐wide survey of reference genes during metamorphism was performed on silkworm <i>Bombyx mori.</i> Twelve genes were chosen as putative reference genes based on a whole genome oligonucleotide microarray normalized by external controls. Then, qRT‐PCR was employed for further validation and selection of potential reference gene candidates. The results were analyzed, and stable genes were selected using geNorm 3.4 and NormFinder software. Finally, considering factors from every aspect, translation initiation factor 4A, translation initiation factor 3 subunit 4, and translation initiation factor 3 subunit 5 (represented by sw22934, swl4876, and swl3956) were selected as reliable internal controls across the examined developmental stages, while cytoplasmic actin (sw22671), the commonly used reference gene in a previous study was shown to vary drastically throughout the examined developmental stages. For future research, we recommend the use of the geometric mean of those three stable reference genes as an accurate normalization factor for data normalization of different developmental stages during metamorphism.</p></abstract></abstractGroup></contentMeta></header></component></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Reference genes identified in the silkworm Bombyx mori during metamorphism based on oligonucleotide microarray and confirmed by qRT‐PCR</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Reference genes identified in the silkworm Bombyx mori during metamorphism based on oligonucleotide microarray and confirmed by qRT‐PCR</title></titleInfo><name type="personal"><namePart type="given">Gen‐Hong</namePart><namePart type="family">Wang</namePart><affiliation>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</affiliation><affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Qing‐You</namePart><namePart type="family">Xia</namePart><affiliation>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</affiliation><affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Dao‐Jun</namePart><namePart type="family">Cheng</namePart><affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Jun</namePart><namePart type="family">Duan</namePart><affiliation>Institute of Agriculture and Life Sciences, Chongqing University, Chongqing, China</affiliation><affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Ping</namePart><namePart type="family">Zhao</namePart><affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Jie</namePart><namePart type="family">Chen</namePart><affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Li</namePart><namePart type="family">Zhu</namePart><affiliation>The Key Sericultural Laboratory of Agricultural Ministry, School of Biotechnology, Southwest University, Chongqing, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="article" displayLabel="article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-6N5SZHKN-D">article</genre><originInfo><publisher>Blackwell Publishing Ltd</publisher><place><placeTerm type="text">Oxford, UK</placeTerm></place><dateIssued encoding="w3cdtf">2008-10</dateIssued><edition>Accepted May 7, 2008</edition><copyrightDate encoding="w3cdtf">2008</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><physicalDescription><extent unit="references">29</extent><extent unit="linksCrossRef">7</extent></physicalDescription><abstract lang="en">Gene expression quantification at mRNA level is very important for post‐genomic studies, as gene expression level is the reflection of the special biological function of the target gene. Methods used for gene expression quantification, such as microarray or quantitative real‐time polymerase chain reaction (qRT‐PCR), require stable expressed reference genes. Thus, finding suitable control genes is essential for gene quantification. In this study, a genome‐wide survey of reference genes during metamorphism was performed on silkworm Bombyx mori. Twelve genes were chosen as putative reference genes based on a whole genome oligonucleotide microarray normalized by external controls. Then, qRT‐PCR was employed for further validation and selection of potential reference gene candidates. The results were analyzed, and stable genes were selected using geNorm 3.4 and NormFinder software. Finally, considering factors from every aspect, translation initiation factor 4A, translation initiation factor 3 subunit 4, and translation initiation factor 3 subunit 5 (represented by sw22934, swl4876, and swl3956) were selected as reliable internal controls across the examined developmental stages, while cytoplasmic actin (sw22671), the commonly used reference gene in a previous study was shown to vary drastically throughout the examined developmental stages. For future research, we recommend the use of the geometric mean of those three stable reference genes as an accurate normalization factor for data normalization of different developmental stages during metamorphism.</abstract><subject lang="en"><genre>keywords</genre><topic>Bombyx mori</topic><topic>microarray</topic><topic>nomalization factor</topic><topic>reference gene</topic><topic>qRT‐PCR</topic></subject><relatedItem type="host"><titleInfo><title>Insect Science</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><identifier type="ISSN">1672-9609</identifier><identifier type="eISSN">1744-7917</identifier><identifier type="DOI">10.1111/(ISSN)1744-7917</identifier><identifier type="PublisherID">INS</identifier><part><date>2008</date><detail type="volume"><caption>vol.</caption><number>15</number></detail><detail type="issue"><caption>no.</caption><number>5</number></detail><extent unit="pages"><start>405</start><end>413</end><total>9</total></extent></part></relatedItem><relatedItem type="references" displayLabel="cit1"><titleInfo><title>Normalization of real‐time quantitative reverse transcription‐PCR data: a model‐based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets</title></titleInfo><name type="personal"><namePart type="given">C.L.</namePart><namePart type="family">Andersen</namePart><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">J.L.</namePart><namePart type="family">Jensen</namePart><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">T.F.</namePart><namePart type="family">Orntoft</namePart><role><roleTerm type="text">author</roleTerm></role></name><genre>journal-article</genre><note type="citation/reference">Andersen, C.L., Jensen, J.L. and Orntoft, T.F. 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Science, 306, 1937–1940.</note><part><date>2004</date><detail type="volume"><caption>vol.</caption><number>306</number></detail><extent unit="pages"><start>1937</start><end>1940</end></extent></part><relatedItem type="host"><titleInfo><title>Science</title></titleInfo><part><date>2004</date><detail type="volume"><caption>vol.</caption><number>306</number></detail><extent unit="pages"><start>1937</start><end>1940</end></extent></part></relatedItem></relatedItem><identifier type="istex">E2A351E4100C5023D8E964D986BEA645E7B0DBCA</identifier><identifier type="ark">ark:/67375/WNG-F0LXHWKL-F</identifier><identifier type="DOI">10.1111/j.1744-7917.2008.00227.x</identifier><identifier type="ArticleID">INS227</identifier><accessCondition type="use and reproduction" contentType="copyright">© 2008 The Authors Journal compilation © Institute of Zoology, Chinese Academy of Sciences</accessCondition><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-L0C46X92-X">wiley</recordContentSource><recordOrigin>Converted from (version ) to MODS version 3.6.</recordOrigin><recordCreationDate encoding="w3cdtf">2019-11-15</recordCreationDate></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/WNG-F0LXHWKL-F/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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