Construction and characterization of two Citrus BAC libraries and identification of clones containing the phytoene synthase gene.
Identifieur interne : 000957 ( PubMed/Curation ); précédent : 000956; suivant : 000958Construction and characterization of two Citrus BAC libraries and identification of clones containing the phytoene synthase gene.
Auteurs : M N R. Baig [République populaire de Chine] ; An Yu ; Wenwu Guo ; Xiuxin DengSource :
- Genome [ 0831-2796 ] ; 2009.
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : Alkyl and Aryl Transferases, DNA, Complementary, DNA, Mitochondrial.
- genetics : Chromosomes, Artificial, Bacterial, Citrus.
- Gene Library, Geranylgeranyl-Diphosphate Geranylgeranyltransferase.
Abstract
Two deep-coverage Bacterial Artificial Chromosome (BAC) libraries of Citrus sinensis (L.) Osbeck 'Cara Cara' navel orange and Citrus reticulata (L.) Blanco 'Egan No. 1' Ponkan mandarin, which belong to the two most important species of the Citrus genus, have been constructed and characterized to facilitate gene cloning and to analyze variety-specific genome composition. The C. sinensis BAC library consists of 36 000 clones with negligible false-positive clones and an estimated average insert size of 126 kb covering ~4.5 x 109 bp and thus providing an 11.8-fold coverage of haploid genome equivalents, whereas the C. reticulata library consists of 21 000 clones also with negligible false-positive clones and an estimated average of 120 kb covering ~2.5 x 109 bp representing a 6.6-fold coverage of haploid genome equivalents. Both libraries were evaluated for contamination with high-copy vector, empty pIndigoBAC536 vector, and organellar DNA sequences. Screening has been performed by Southern hybridization of BAC filters, which results in <0.5% chloroplast DNA contamination and no mitochondrial DNA contamination in both libraries. Eight and five positive clones harboring the gene encoding Phytoene synthase (Psy (EC 2.5.1.32)) were identified from the C. sinensis and C. reticulata libraries, respectively, using the filter hybridization procedure. These results suggest that the two BAC libraries are useful tools for the isolation of functional genes and advanced genomics research in the two important species C. sinensis and C. reticulata. Resources, high-density filters, individual clones, and whole libraries are available for public distribution and are accessible at the National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University.
DOI: 10.1139/g09-017
PubMed: 19448728
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Baig</name><affiliation wicri:level="1"><nlm:affiliation>National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070</wicri:regionArea></affiliation></author><author><name sortKey="Yu, An" sort="Yu, An" uniqKey="Yu A" first="An" last="Yu">An Yu</name></author><author><name sortKey="Guo, Wenwu" sort="Guo, Wenwu" uniqKey="Guo W" first="Wenwu" last="Guo">Wenwu Guo</name></author><author><name sortKey="Deng, Xiuxin" sort="Deng, Xiuxin" uniqKey="Deng X" first="Xiuxin" last="Deng">Xiuxin Deng</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2009">2009</date><idno type="RBID">pubmed:19448728</idno><idno type="pmid">19448728</idno><idno type="doi">10.1139/g09-017</idno><idno type="wicri:Area/PubMed/Corpus">000957</idno><idno type="wicri:Area/PubMed/Curation">000957</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Construction and characterization of two Citrus BAC libraries and identification of clones containing the phytoene synthase gene.</title><author><name sortKey="Baig, M N R" sort="Baig, M N R" uniqKey="Baig M" first="M N R" last="Baig">M N R. 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Osbeck 'Cara Cara' navel orange and Citrus reticulata (L.) Blanco 'Egan No. 1' Ponkan mandarin, which belong to the two most important species of the Citrus genus, have been constructed and characterized to facilitate gene cloning and to analyze variety-specific genome composition. The C. sinensis BAC library consists of 36 000 clones with negligible false-positive clones and an estimated average insert size of 126 kb covering ~4.5 x 109 bp and thus providing an 11.8-fold coverage of haploid genome equivalents, whereas the C. reticulata library consists of 21 000 clones also with negligible false-positive clones and an estimated average of 120 kb covering ~2.5 x 109 bp representing a 6.6-fold coverage of haploid genome equivalents. Both libraries were evaluated for contamination with high-copy vector, empty pIndigoBAC536 vector, and organellar DNA sequences. Screening has been performed by Southern hybridization of BAC filters, which results in <0.5% chloroplast DNA contamination and no mitochondrial DNA contamination in both libraries. Eight and five positive clones harboring the gene encoding Phytoene synthase (Psy (EC 2.5.1.32)) were identified from the C. sinensis and C. reticulata libraries, respectively, using the filter hybridization procedure. These results suggest that the two BAC libraries are useful tools for the isolation of functional genes and advanced genomics research in the two important species C. sinensis and C. reticulata. Resources, high-density filters, individual clones, and whole libraries are available for public distribution and are accessible at the National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">19448728</PMID><DateCreated><Year>2009</Year><Month>5</Month><Day>18</Day></DateCreated><DateCompleted><Year>2009</Year><Month>08</Month><Day>14</Day></DateCompleted><DateRevised><Year>2016</Year><Month>10</Month><Day>20</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0831-2796</ISSN><JournalIssue CitedMedium="Print"><Volume>52</Volume><Issue>5</Issue><PubDate><Year>2009</Year><Month>May</Month></PubDate></JournalIssue><Title>Genome</Title><ISOAbbreviation>Genome</ISOAbbreviation></Journal><ArticleTitle>Construction and characterization of two Citrus BAC libraries and identification of clones containing the phytoene synthase gene.</ArticleTitle><Pagination><MedlinePgn>484-9</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1139/g09-017</ELocationID><Abstract><AbstractText>Two deep-coverage Bacterial Artificial Chromosome (BAC) libraries of Citrus sinensis (L.) Osbeck 'Cara Cara' navel orange and Citrus reticulata (L.) Blanco 'Egan No. 1' Ponkan mandarin, which belong to the two most important species of the Citrus genus, have been constructed and characterized to facilitate gene cloning and to analyze variety-specific genome composition. The C. sinensis BAC library consists of 36 000 clones with negligible false-positive clones and an estimated average insert size of 126 kb covering ~4.5 x 109 bp and thus providing an 11.8-fold coverage of haploid genome equivalents, whereas the C. reticulata library consists of 21 000 clones also with negligible false-positive clones and an estimated average of 120 kb covering ~2.5 x 109 bp representing a 6.6-fold coverage of haploid genome equivalents. Both libraries were evaluated for contamination with high-copy vector, empty pIndigoBAC536 vector, and organellar DNA sequences. Screening has been performed by Southern hybridization of BAC filters, which results in <0.5% chloroplast DNA contamination and no mitochondrial DNA contamination in both libraries. Eight and five positive clones harboring the gene encoding Phytoene synthase (Psy (EC 2.5.1.32)) were identified from the C. sinensis and C. reticulata libraries, respectively, using the filter hybridization procedure. These results suggest that the two BAC libraries are useful tools for the isolation of functional genes and advanced genomics research in the two important species C. sinensis and C. reticulata. 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