Systematic Identification of Essential Genes by in vitro mariner Mutagenesis
Although the complete DNA sequences of several microbial genomes are now available, nearly 40% of the putative genes lack identifiable functions. Comprehensive screens and selections for identifying functional classes of genes are needed to convert sequence data into meaningful biological informatio...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1998-07, Vol.95 (15), p.8927-8932 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
| container_volume | 95 |
| creator | Akerley, Brian J. Rubin, Eric J. Camilli, Andrew Lampe, David J. Robertson, Hugh M. Mekalanos, John J. |
| description | Although the complete DNA sequences of several microbial genomes are now available, nearly 40% of the putative genes lack identifiable functions. Comprehensive screens and selections for identifying functional classes of genes are needed to convert sequence data into meaningful biological information. One particularly significant group of bacterial genes consists of those that are essential for growth or viability. Here, we describe a simple system for performing transposon mutagenesis on naturally transformable organisms along with a technique to rapidly identify essential or conditionally essential DNA segments. We show the general utility of this approach by applying it to two human pathogens, Haemophilus influenzae and Streptococcus pneumoniae, in which we detected known essential genes and assigned essentiality to several ORFs of unknown function. |
| doi_str_mv | 10.1073/pnas.95.15.8927 |
| format | Article |
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Comprehensive screens and selections for identifying functional classes of genes are needed to convert sequence data into meaningful biological information. One particularly significant group of bacterial genes consists of those that are essential for growth or viability. Here, we describe a simple system for performing transposon mutagenesis on naturally transformable organisms along with a technique to rapidly identify essential or conditionally essential DNA segments. 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Comprehensive screens and selections for identifying functional classes of genes are needed to convert sequence data into meaningful biological information. One particularly significant group of bacterial genes consists of those that are essential for growth or viability. Here, we describe a simple system for performing transposon mutagenesis on naturally transformable organisms along with a technique to rapidly identify essential or conditionally essential DNA segments. We show the general utility of this approach by applying it to two human pathogens, Haemophilus influenzae and Streptococcus pneumoniae, in which we detected known essential genes and assigned essentiality to several ORFs of unknown function.</description><subject>Bacteria</subject><subject>Base Sequence</subject><subject>Biological Sciences</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA Primers</subject><subject>DNA Transposable Elements</subject><subject>DNA-Binding Proteins</subject><subject>Essential genes</subject><subject>Gels</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Genetic transposition</subject><subject>Haemophilus influenzae - genetics</subject><subject>Mutagenesis</subject><subject>Mutation</subject><subject>Open Reading Frames</subject><subject>Polymerase chain reaction</subject><subject>Product category rules</subject><subject>Streptococcus pneumoniae - genetics</subject><subject>Transposases</subject><subject>Transposons</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kcFrFDEUxoModa2eBUEJHvQ027xkkkmgFym1FlY8qOeQSTM1y2yyJpni_vdm2GVQD57Cy_f7Ht_jQ-glkDWQjl3sg8lrxdfA11LR7hFaAVHQiFaRx2hFCO0a2dL2KXqW85YQorgkZ-hMiQ46CSu0-XrIxe1M8Rbf3rlQ_OBtnWLAccDXOc9fZsQ3LriM-wP2AT_4kiLemeSDS_jzVMz9rPr8HD0ZzJjdi9N7jr5_vP529anZfLm5vfqwaSwHWRrVcnBEuL4FxigxordGDRZqKM5tz4CCFFYxSdtBdWzgrTM9sbbeYR1Ywc7R5XHvfup37s7WjMmMep98DXXQ0Xj9txL8D30fHzQF6FS1vzvZU_w5uVz0zmfrxtEEF6esQXDKWUsq-PYfcBunFOppmhJgnSBcVujiCNkUc05uWHIA0XNHeu5IK66B67mj6nj9Z_yFP5VS9fcnfTYu6rJAD9M4FverVPLNf8kKvDoC21xiWoiWS0HZb9puryk</recordid><startdate>19980721</startdate><enddate>19980721</enddate><creator>Akerley, Brian J.</creator><creator>Rubin, Eric J.</creator><creator>Camilli, Andrew</creator><creator>Lampe, David J.</creator><creator>Robertson, Hugh M.</creator><creator>Mekalanos, John J.</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><general>National Academy of Sciences</general><general>The National Academy of Sciences</general><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>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>19980721</creationdate><title>Systematic Identification of Essential Genes by in vitro mariner Mutagenesis</title><author>Akerley, Brian J. ; Rubin, Eric J. ; Camilli, Andrew ; Lampe, David J. ; Robertson, Hugh M. ; Mekalanos, John J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c518t-9451e06eb413320a6bca9fc196755cb312186c93824f973f54eab0cc424ce1c63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>Bacteria</topic><topic>Base Sequence</topic><topic>Biological Sciences</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA Primers</topic><topic>DNA Transposable Elements</topic><topic>DNA-Binding Proteins</topic><topic>Essential genes</topic><topic>Gels</topic><topic>Genes</topic><topic>Genes, Bacterial</topic><topic>Genetic transposition</topic><topic>Haemophilus influenzae - genetics</topic><topic>Mutagenesis</topic><topic>Mutation</topic><topic>Open Reading Frames</topic><topic>Polymerase chain reaction</topic><topic>Product category rules</topic><topic>Streptococcus pneumoniae - genetics</topic><topic>Transposases</topic><topic>Transposons</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Akerley, Brian J.</creatorcontrib><creatorcontrib>Rubin, Eric J.</creatorcontrib><creatorcontrib>Camilli, Andrew</creatorcontrib><creatorcontrib>Lampe, David J.</creatorcontrib><creatorcontrib>Robertson, Hugh M.</creatorcontrib><creatorcontrib>Mekalanos, John J.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Akerley, Brian J.</au><au>Rubin, Eric J.</au><au>Camilli, Andrew</au><au>Lampe, David J.</au><au>Robertson, Hugh M.</au><au>Mekalanos, John J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Systematic Identification of Essential Genes by in vitro mariner Mutagenesis</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1998-07-21</date><risdate>1998</risdate><volume>95</volume><issue>15</issue><spage>8927</spage><epage>8932</epage><pages>8927-8932</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Although the complete DNA sequences of several microbial genomes are now available, nearly 40% of the putative genes lack identifiable functions. 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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1998-07, Vol.95 (15), p.8927-8932 |
| issn | 0027-8424 1091-6490 |
| language | eng |
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| source | Jstor Complete Legacy; MEDLINE; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | Bacteria Base Sequence Biological Sciences Deoxyribonucleic acid DNA DNA Primers DNA Transposable Elements DNA-Binding Proteins Essential genes Gels Genes Genes, Bacterial Genetic transposition Haemophilus influenzae - genetics Mutagenesis Mutation Open Reading Frames Polymerase chain reaction Product category rules Streptococcus pneumoniae - genetics Transposases Transposons |
| title | Systematic Identification of Essential Genes by in vitro mariner Mutagenesis |
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