[Establishment of mammalian cell lines for constitutive expression of influenza virus matrix protein 2].
Identifieur interne : 000805 ( Main/Exploration ); précédent : 000804; suivant : 000806[Establishment of mammalian cell lines for constitutive expression of influenza virus matrix protein 2].
Auteurs : Ai-Jun Chen [République populaire de Chine] ; Jian-Qiang Guo ; Li-Hong Yao ; Ji-Yong Yin ; Zhi-Qing ZhangSource :
- Bing du xue bao = Chinese journal of virology [ 1000-8721 ] ; 2013.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , biosynthesis : Recombinant Proteins, Viral Matrix Proteins.
- chemistry : Influenza A Virus, H2N2 Subtype.
- chemical , genetics : Viral Matrix Proteins.
- Animals, CHO Cells, Cell Culture Techniques, Cell Line, Cricetinae, Cricetulus.
Abstract
To establish a mammalian cell line for stable expression of the matrix protein 2 (M2) of influenza virus type A. M2 gene was amplified by PCR from the influenza virus strain A/PR/8/34. The PCR product was cloned into eukaryotic expression vector pcDNA5/FRT. After identification with restriction enzyme digestion, the plasmid was co-transfected with plasmid pOG44 which expressed Flp in Flp-In-CHO cells. The target gene was integrated into chromosome of CHO cells by homologous recombination in vivo. Recombinant CHO-M2 cell lines were selected for hygromycin B resistance. A total of 15 recombinant cell strains with high expression of M2 protein were screened by hygromycin, and the expression of M2 protein was determined by IFA and Western blot. After subculturing for 10 passages, the presence of M2 gene in the CHO-M2 cells was confirmed by PCR, and the expression of M2 protein were proved by IFA and Western blot. We successfully constructed a mammalian cell line which stably expressed M2 protein of influenza virus type A. The cell line will be useful for studies on function of M2 protein and provide tools for novel influenza virus vaccine development.
PubMed: 23547373
Affiliations:
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J" first="Jian-Qiang" last="Guo">Jian-Qiang Guo</name></author><author><name sortKey="Yao, Li Hong" sort="Yao, Li Hong" uniqKey="Yao L" first="Li-Hong" last="Yao">Li-Hong Yao</name></author><author><name sortKey="Yin, Ji Yong" sort="Yin, Ji Yong" uniqKey="Yin J" first="Ji-Yong" last="Yin">Ji-Yong Yin</name></author><author><name sortKey="Zhang, Zhi Qing" sort="Zhang, Zhi Qing" uniqKey="Zhang Z" first="Zhi-Qing" last="Zhang">Zhi-Qing Zhang</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2013">2013</date><idno type="RBID">pubmed:23547373</idno><idno type="pmid">23547373</idno><idno type="wicri:Area/PubMed/Corpus">000126</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000126</idno><idno type="wicri:Area/PubMed/Curation">000126</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000126</idno><idno type="wicri:Area/PubMed/Checkpoint">000104</idno><idno 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China. chenajsun@yahoo.com.cn</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Molecular Virology and Genetic Engineering Laboratory, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Guo, Jian Qiang" sort="Guo, Jian Qiang" uniqKey="Guo J" first="Jian-Qiang" last="Guo">Jian-Qiang Guo</name></author><author><name sortKey="Yao, Li Hong" sort="Yao, Li Hong" uniqKey="Yao L" first="Li-Hong" last="Yao">Li-Hong Yao</name></author><author><name sortKey="Yin, Ji Yong" sort="Yin, Ji Yong" uniqKey="Yin J" first="Ji-Yong" last="Yin">Ji-Yong Yin</name></author><author><name sortKey="Zhang, Zhi Qing" sort="Zhang, Zhi Qing" uniqKey="Zhang Z" first="Zhi-Qing" last="Zhang">Zhi-Qing Zhang</name></author></analytic><series><title level="j">Bing du xue bao = Chinese journal of virology</title><idno type="ISSN">1000-8721</idno><imprint><date when="2013" type="published">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>CHO Cells</term><term>Cell Culture Techniques</term><term>Cell Line</term><term>Cricetinae</term><term>Cricetulus</term><term>Influenza A Virus, H2N2 Subtype (chemistry)</term><term>Recombinant Proteins (biosynthesis)</term><term>Viral Matrix Proteins (biosynthesis)</term><term>Viral Matrix Proteins (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Cellules CHO</term><term>Cricetinae</term><term>Cricetulus</term><term>Lignée cellulaire</term><term>Protéines de la matrice virale (biosynthèse)</term><term>Protéines de la matrice virale (génétique)</term><term>Protéines recombinantes (biosynthèse)</term><term>Sous-type H2N2 du virus de la grippe A ()</term><term>Techniques de culture cellulaire</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Recombinant Proteins</term><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Protéines de la matrice virale</term><term>Protéines recombinantes</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Influenza A Virus, H2N2 Subtype</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines de la matrice virale</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>CHO Cells</term><term>Cell Culture Techniques</term><term>Cell Line</term><term>Cricetinae</term><term>Cricetulus</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules CHO</term><term>Cricetinae</term><term>Cricetulus</term><term>Lignée cellulaire</term><term>Sous-type H2N2 du virus de la grippe A</term><term>Techniques de culture cellulaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">To establish a mammalian cell line for stable expression of the matrix protein 2 (M2) of influenza virus type A. M2 gene was amplified by PCR from the influenza virus strain A/PR/8/34. The PCR product was cloned into eukaryotic expression vector pcDNA5/FRT. After identification with restriction enzyme digestion, the plasmid was co-transfected with plasmid pOG44 which expressed Flp in Flp-In-CHO cells. The target gene was integrated into chromosome of CHO cells by homologous recombination in vivo. Recombinant CHO-M2 cell lines were selected for hygromycin B resistance. A total of 15 recombinant cell strains with high expression of M2 protein were screened by hygromycin, and the expression of M2 protein was determined by IFA and Western blot. After subculturing for 10 passages, the presence of M2 gene in the CHO-M2 cells was confirmed by PCR, and the expression of M2 protein were proved by IFA and Western blot. We successfully constructed a mammalian cell line which stably expressed M2 protein of influenza virus type A. The cell line will be useful for studies on function of M2 protein and provide tools for novel influenza virus vaccine development.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><settlement><li>Pékin</li></settlement></list><tree><noCountry><name sortKey="Guo, Jian Qiang" sort="Guo, Jian Qiang" uniqKey="Guo J" first="Jian-Qiang" last="Guo">Jian-Qiang Guo</name><name sortKey="Yao, Li Hong" sort="Yao, Li Hong" uniqKey="Yao L" first="Li-Hong" last="Yao">Li-Hong Yao</name><name sortKey="Yin, Ji Yong" sort="Yin, Ji Yong" uniqKey="Yin J" first="Ji-Yong" last="Yin">Ji-Yong Yin</name><name sortKey="Zhang, Zhi Qing" sort="Zhang, Zhi Qing" uniqKey="Zhang Z" first="Zhi-Qing" last="Zhang">Zhi-Qing Zhang</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Chen, Ai Jun" sort="Chen, Ai Jun" uniqKey="Chen A" first="Ai-Jun" last="Chen">Ai-Jun Chen</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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