A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda
Two genomic tools for the study of Lepidoptera and the holocentric structure of their chromosomes are presented in this paper. A bacterial artificial chromosome (BAC) library was constructed using nuclear DNA partially digested with HindIII from eggs of Spodoptera frugiperda. The library contains a total of 36 864 clones with an average insert size of 125 kb, which corresponds to approximately 11.5 genome equivalents. Hybridization screening of the library was performed with eight single-copy genes, giving an average hit of 10 clones per marker gene. Colinearity between the genome and BACs was demonstrated at the triose phosphate isomerase (tpi) locus. Probing of the library with a PCR fragment internal to the 18S ribosomal gene allowed an estimation of the rDNA locus size close to 115 repeats per haploid genome. A new vector (pBAC3.6eGFP) for transient transfection into S. frugiperda cell lines has been constructed. It is based on the BAC vector, pBAC3.6e, in which a gene encoding GFP was inserted under the control of the densovirus P9 promoter. This vector has the advantage to accommodate large genomic inserts and to be transfected in a large lepidopteran host range. It was used to construct a second BAC library from Sf9 cell nuclear DNA in order to allow a comparison between somatic and cell line genome organization.
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Subjects: | F30 - Génétique et amélioration des plantes, Spodoptera frugiperda, insecte nuisible, biotechnologie animale, vecteur génétique, chromosome, banque de gènes, adn, adn ribosomal, http://aims.fao.org/aos/agrovoc/c_25452, http://aims.fao.org/aos/agrovoc/c_5729, http://aims.fao.org/aos/agrovoc/c_27450, http://aims.fao.org/aos/agrovoc/c_27532, http://aims.fao.org/aos/agrovoc/c_1598, http://aims.fao.org/aos/agrovoc/c_11116, http://aims.fao.org/aos/agrovoc/c_2347, http://aims.fao.org/aos/agrovoc/c_1354098656894, |
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dig-cirad-fr-5194562024-01-28T12:21:18Z http://agritrop.cirad.fr/519456/ http://agritrop.cirad.fr/519456/ A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda. D'Alençon Emmanuelle, Piffanelli Pietro, Volkoff Anne-Nathalie, Sabau Xavier, Gimenez Sylvie, Rocher Janick, Cérutti Pierre, Fournier Philippe. 2004. Insect Biochemistry and Molecular Biology, 34 (4) : 331-341.https://doi.org/10.1016/j.ibmb.2003.12.004 <https://doi.org/10.1016/j.ibmb.2003.12.004> A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda D'Alençon, Emmanuelle Piffanelli, Pietro Volkoff, Anne-Nathalie Sabau, Xavier Gimenez, Sylvie Rocher, Janick Cérutti, Pierre Fournier, Philippe eng 2004 Insect Biochemistry and Molecular Biology F30 - Génétique et amélioration des plantes Spodoptera frugiperda insecte nuisible biotechnologie animale vecteur génétique chromosome banque de gènes adn adn ribosomal http://aims.fao.org/aos/agrovoc/c_25452 http://aims.fao.org/aos/agrovoc/c_5729 http://aims.fao.org/aos/agrovoc/c_27450 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_1598 http://aims.fao.org/aos/agrovoc/c_11116 http://aims.fao.org/aos/agrovoc/c_2347 http://aims.fao.org/aos/agrovoc/c_1354098656894 Two genomic tools for the study of Lepidoptera and the holocentric structure of their chromosomes are presented in this paper. A bacterial artificial chromosome (BAC) library was constructed using nuclear DNA partially digested with HindIII from eggs of Spodoptera frugiperda. The library contains a total of 36 864 clones with an average insert size of 125 kb, which corresponds to approximately 11.5 genome equivalents. Hybridization screening of the library was performed with eight single-copy genes, giving an average hit of 10 clones per marker gene. Colinearity between the genome and BACs was demonstrated at the triose phosphate isomerase (tpi) locus. Probing of the library with a PCR fragment internal to the 18S ribosomal gene allowed an estimation of the rDNA locus size close to 115 repeats per haploid genome. A new vector (pBAC3.6eGFP) for transient transfection into S. frugiperda cell lines has been constructed. It is based on the BAC vector, pBAC3.6e, in which a gene encoding GFP was inserted under the control of the densovirus P9 promoter. This vector has the advantage to accommodate large genomic inserts and to be transfected in a large lepidopteran host range. It was used to construct a second BAC library from Sf9 cell nuclear DNA in order to allow a comparison between somatic and cell line genome organization. article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/519456/1/519456.pdf text Cirad license info:eu-repo/semantics/restrictedAccess https://agritrop.cirad.fr/mention_legale.html https://doi.org/10.1016/j.ibmb.2003.12.004 10.1016/j.ibmb.2003.12.004 http://catalogue-bibliotheques.cirad.fr/cgi-bin/koha/opac-detail.pl?biblionumber=181027 info:eu-repo/semantics/altIdentifier/doi/10.1016/j.ibmb.2003.12.004 info:eu-repo/semantics/altIdentifier/purl/https://doi.org/10.1016/j.ibmb.2003.12.004 |
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F30 - Génétique et amélioration des plantes Spodoptera frugiperda insecte nuisible biotechnologie animale vecteur génétique chromosome banque de gènes adn adn ribosomal http://aims.fao.org/aos/agrovoc/c_25452 http://aims.fao.org/aos/agrovoc/c_5729 http://aims.fao.org/aos/agrovoc/c_27450 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_1598 http://aims.fao.org/aos/agrovoc/c_11116 http://aims.fao.org/aos/agrovoc/c_2347 http://aims.fao.org/aos/agrovoc/c_1354098656894 F30 - Génétique et amélioration des plantes Spodoptera frugiperda insecte nuisible biotechnologie animale vecteur génétique chromosome banque de gènes adn adn ribosomal http://aims.fao.org/aos/agrovoc/c_25452 http://aims.fao.org/aos/agrovoc/c_5729 http://aims.fao.org/aos/agrovoc/c_27450 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_1598 http://aims.fao.org/aos/agrovoc/c_11116 http://aims.fao.org/aos/agrovoc/c_2347 http://aims.fao.org/aos/agrovoc/c_1354098656894 |
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F30 - Génétique et amélioration des plantes Spodoptera frugiperda insecte nuisible biotechnologie animale vecteur génétique chromosome banque de gènes adn adn ribosomal http://aims.fao.org/aos/agrovoc/c_25452 http://aims.fao.org/aos/agrovoc/c_5729 http://aims.fao.org/aos/agrovoc/c_27450 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_1598 http://aims.fao.org/aos/agrovoc/c_11116 http://aims.fao.org/aos/agrovoc/c_2347 http://aims.fao.org/aos/agrovoc/c_1354098656894 F30 - Génétique et amélioration des plantes Spodoptera frugiperda insecte nuisible biotechnologie animale vecteur génétique chromosome banque de gènes adn adn ribosomal http://aims.fao.org/aos/agrovoc/c_25452 http://aims.fao.org/aos/agrovoc/c_5729 http://aims.fao.org/aos/agrovoc/c_27450 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_1598 http://aims.fao.org/aos/agrovoc/c_11116 http://aims.fao.org/aos/agrovoc/c_2347 http://aims.fao.org/aos/agrovoc/c_1354098656894 D'Alençon, Emmanuelle Piffanelli, Pietro Volkoff, Anne-Nathalie Sabau, Xavier Gimenez, Sylvie Rocher, Janick Cérutti, Pierre Fournier, Philippe A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda |
description |
Two genomic tools for the study of Lepidoptera and the holocentric structure of their chromosomes are presented in this paper. A bacterial artificial chromosome (BAC) library was constructed using nuclear DNA partially digested with HindIII from eggs of Spodoptera frugiperda. The library contains a total of 36 864 clones with an average insert size of 125 kb, which corresponds to approximately 11.5 genome equivalents. Hybridization screening of the library was performed with eight single-copy genes, giving an average hit of 10 clones per marker gene. Colinearity between the genome and BACs was demonstrated at the triose phosphate isomerase (tpi) locus. Probing of the library with a PCR fragment internal to the 18S ribosomal gene allowed an estimation of the rDNA locus size close to 115 repeats per haploid genome. A new vector (pBAC3.6eGFP) for transient transfection into S. frugiperda cell lines has been constructed. It is based on the BAC vector, pBAC3.6e, in which a gene encoding GFP was inserted under the control of the densovirus P9 promoter. This vector has the advantage to accommodate large genomic inserts and to be transfected in a large lepidopteran host range. It was used to construct a second BAC library from Sf9 cell nuclear DNA in order to allow a comparison between somatic and cell line genome organization. |
format |
article |
topic_facet |
F30 - Génétique et amélioration des plantes Spodoptera frugiperda insecte nuisible biotechnologie animale vecteur génétique chromosome banque de gènes adn adn ribosomal http://aims.fao.org/aos/agrovoc/c_25452 http://aims.fao.org/aos/agrovoc/c_5729 http://aims.fao.org/aos/agrovoc/c_27450 http://aims.fao.org/aos/agrovoc/c_27532 http://aims.fao.org/aos/agrovoc/c_1598 http://aims.fao.org/aos/agrovoc/c_11116 http://aims.fao.org/aos/agrovoc/c_2347 http://aims.fao.org/aos/agrovoc/c_1354098656894 |
author |
D'Alençon, Emmanuelle Piffanelli, Pietro Volkoff, Anne-Nathalie Sabau, Xavier Gimenez, Sylvie Rocher, Janick Cérutti, Pierre Fournier, Philippe |
author_facet |
D'Alençon, Emmanuelle Piffanelli, Pietro Volkoff, Anne-Nathalie Sabau, Xavier Gimenez, Sylvie Rocher, Janick Cérutti, Pierre Fournier, Philippe |
author_sort |
D'Alençon, Emmanuelle |
title |
A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda |
title_short |
A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda |
title_full |
A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda |
title_fullStr |
A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda |
title_full_unstemmed |
A genomic BAC library and a new BAC-GFP vector to study the holocentric pest Spodoptera frugiperda |
title_sort |
genomic bac library and a new bac-gfp vector to study the holocentric pest spodoptera frugiperda |
url |
http://agritrop.cirad.fr/519456/ http://agritrop.cirad.fr/519456/1/519456.pdf |
work_keys_str_mv |
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