In vivo selection and characterization of internal deletions in the lamB::lacZ gene fusion
Strain Pop3299 contains the lamB:: lacZ42-12 gene fusion that encodes a hybrid protein that is efficiently exported to the cellular envelope of Escherichia coli. As a result of this efficient export, this strain is killed by the inducer maltose and unable to grow on minimal media supplemented with l...
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| Veröffentlicht in: | Gene 1987, Vol.52 (2), p.165-173 |
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| container_title | Gene |
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| creator | Benson, Spencer A. Bremer, Erhard |
| description | Strain Pop3299 contains the lamB:: lacZ42-12 gene fusion that encodes a hybrid protein that is efficiently exported to the cellular envelope of
Escherichia coli. As a result of this efficient export, this strain is killed by the inducer maltose and unable to grow on minimal media supplemented with lactose. In an attempt to isolate mutants in which export of the hybrid protein is altered, we selected Lac
+ mutants of strain Pop3299 on lactose tetrazolium media. Unlike mutants previously isolated on lactose minimal media, all the mutants we obtained carried large deletions within the
lamB:: lacZ gene fusion. Thus, it appears that the type of selection employed affects the type of mutations obtained. We have analyzed the nucleotide sequences of representative mutants, and demonstrate a correlation between the deletion size and the export-related maltose and lactose phenotypes. In addition, we demonstrate that the deletions do not appear to arise from regions of micro-homology. |
| doi_str_mv | 10.1016/0378-1119(87)90043-6 |
| format | Article |
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Escherichia coli. As a result of this efficient export, this strain is killed by the inducer maltose and unable to grow on minimal media supplemented with lactose. In an attempt to isolate mutants in which export of the hybrid protein is altered, we selected Lac
+ mutants of strain Pop3299 on lactose tetrazolium media. Unlike mutants previously isolated on lactose minimal media, all the mutants we obtained carried large deletions within the
lamB:: lacZ gene fusion. Thus, it appears that the type of selection employed affects the type of mutations obtained. We have analyzed the nucleotide sequences of representative mutants, and demonstrate a correlation between the deletion size and the export-related maltose and lactose phenotypes. In addition, we demonstrate that the deletions do not appear to arise from regions of micro-homology.</description><identifier>ISSN: 0378-1119</identifier><identifier>EISSN: 1879-0038</identifier><identifier>DOI: 10.1016/0378-1119(87)90043-6</identifier><identifier>PMID: 3038681</identifier><identifier>CODEN: GENED6</identifier><language>eng</language><publisher>Lausanne: Elsevier B.V</publisher><subject>Bacterial Outer Membrane Proteins - genetics ; Bacterial Proteins - genetics ; Bacteriology ; Bacteriophage lambda - metabolism ; Base Sequence ; Biological and medical sciences ; Chromosome Deletion ; E. coli ; Escherichia coli ; Escherichia coli - genetics ; Fundamental and applied biological sciences. Psychology ; Genes ; Genes, Bacterial ; lactose utilization ; Microbiology ; Mutation ; nucleotide sequences ; Permeability, membrane transport, intracellular transport ; phage Mu ; Porins ; protein export ; Receptors, Virus - genetics ; Recombinant DNA ; spontaneous mutants</subject><ispartof>Gene, 1987, Vol.52 (2), p.165-173</ispartof><rights>1987</rights><rights>1988 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c332t-1e19cf5068d4fa3d846a845ab646cda50447299b72ca58094741551234d8c8c43</citedby><cites>FETCH-LOGICAL-c332t-1e19cf5068d4fa3d846a845ab646cda50447299b72ca58094741551234d8c8c43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/0378-1119(87)90043-6$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,4024,27923,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7442648$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3038681$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Benson, Spencer A.</creatorcontrib><creatorcontrib>Bremer, Erhard</creatorcontrib><title>In vivo selection and characterization of internal deletions in the lamB::lacZ gene fusion</title><title>Gene</title><addtitle>Gene</addtitle><description>Strain Pop3299 contains the lamB:: lacZ42-12 gene fusion that encodes a hybrid protein that is efficiently exported to the cellular envelope of
Escherichia coli. As a result of this efficient export, this strain is killed by the inducer maltose and unable to grow on minimal media supplemented with lactose. In an attempt to isolate mutants in which export of the hybrid protein is altered, we selected Lac
+ mutants of strain Pop3299 on lactose tetrazolium media. Unlike mutants previously isolated on lactose minimal media, all the mutants we obtained carried large deletions within the
lamB:: lacZ gene fusion. Thus, it appears that the type of selection employed affects the type of mutations obtained. We have analyzed the nucleotide sequences of representative mutants, and demonstrate a correlation between the deletion size and the export-related maltose and lactose phenotypes. In addition, we demonstrate that the deletions do not appear to arise from regions of micro-homology.</description><subject>Bacterial Outer Membrane Proteins - genetics</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacteriology</subject><subject>Bacteriophage lambda - metabolism</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Chromosome Deletion</subject><subject>E. coli</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>lactose utilization</subject><subject>Microbiology</subject><subject>Mutation</subject><subject>nucleotide sequences</subject><subject>Permeability, membrane transport, intracellular transport</subject><subject>phage Mu</subject><subject>Porins</subject><subject>protein export</subject><subject>Receptors, Virus - genetics</subject><subject>Recombinant DNA</subject><subject>spontaneous mutants</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9rFDEUx4ModVv9DxRyENHDaDL53UNBS9VCwYteeglvkzc2MpupyeyC_vVmusseNZeQ9_28x-MTQl5w9o4zrt8zYWzHOXdvrHnrGJOi04_IilvjOsaEfUxWR-QpOa31J2tHqf6EnIiWa8tX5PY6013aTbTiiGFOU6aQIw13UCDMWNIfeChOA025vTOMNDZ0KdZWovMd0hE2H8_PRwi39AdmpMO2tvgZeTLAWPH54T4j3z9dfbv80t18_Xx9-eGmC0L0c8eRuzAopm2UA4hopQYrFay11CGCYlKa3rm16QMoy5w0kivFeyGjDTZIcUZe7-fel-nXFuvsN6kGHEfIOG2rN0Y5I7j7L8il0UapBZR7MJSp1oKDvy9pA-W358wv7v0i1i9ivTX-wb3Xre3lYf52vcF4bDrIbvmrQw41wDgUyCHVI2ak7LW0DbvYY9ik7RIWX0PCHDCm0r7Ixyn9e4-_leeehQ</recordid><startdate>1987</startdate><enddate>1987</enddate><creator>Benson, Spencer A.</creator><creator>Bremer, Erhard</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><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>7QL</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>1987</creationdate><title>In vivo selection and characterization of internal deletions in the lamB::lacZ gene fusion</title><author>Benson, Spencer A. ; Bremer, Erhard</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c332t-1e19cf5068d4fa3d846a845ab646cda50447299b72ca58094741551234d8c8c43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>Bacterial Outer Membrane Proteins - genetics</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacteriology</topic><topic>Bacteriophage lambda - metabolism</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Chromosome Deletion</topic><topic>E. coli</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes</topic><topic>Genes, Bacterial</topic><topic>lactose utilization</topic><topic>Microbiology</topic><topic>Mutation</topic><topic>nucleotide sequences</topic><topic>Permeability, membrane transport, intracellular transport</topic><topic>phage Mu</topic><topic>Porins</topic><topic>protein export</topic><topic>Receptors, Virus - genetics</topic><topic>Recombinant DNA</topic><topic>spontaneous mutants</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Benson, Spencer A.</creatorcontrib><creatorcontrib>Bremer, Erhard</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Benson, Spencer A.</au><au>Bremer, Erhard</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In vivo selection and characterization of internal deletions in the lamB::lacZ gene fusion</atitle><jtitle>Gene</jtitle><addtitle>Gene</addtitle><date>1987</date><risdate>1987</risdate><volume>52</volume><issue>2</issue><spage>165</spage><epage>173</epage><pages>165-173</pages><issn>0378-1119</issn><eissn>1879-0038</eissn><coden>GENED6</coden><abstract>Strain Pop3299 contains the lamB:: lacZ42-12 gene fusion that encodes a hybrid protein that is efficiently exported to the cellular envelope of
Escherichia coli. As a result of this efficient export, this strain is killed by the inducer maltose and unable to grow on minimal media supplemented with lactose. In an attempt to isolate mutants in which export of the hybrid protein is altered, we selected Lac
+ mutants of strain Pop3299 on lactose tetrazolium media. Unlike mutants previously isolated on lactose minimal media, all the mutants we obtained carried large deletions within the
lamB:: lacZ gene fusion. Thus, it appears that the type of selection employed affects the type of mutations obtained. We have analyzed the nucleotide sequences of representative mutants, and demonstrate a correlation between the deletion size and the export-related maltose and lactose phenotypes. In addition, we demonstrate that the deletions do not appear to arise from regions of micro-homology.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>3038681</pmid><doi>10.1016/0378-1119(87)90043-6</doi><tpages>9</tpages></addata></record> |
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| ispartof | Gene, 1987, Vol.52 (2), p.165-173 |
| issn | 0378-1119 1879-0038 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Bacterial Outer Membrane Proteins - genetics Bacterial Proteins - genetics Bacteriology Bacteriophage lambda - metabolism Base Sequence Biological and medical sciences Chromosome Deletion E. coli Escherichia coli Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Genes Genes, Bacterial lactose utilization Microbiology Mutation nucleotide sequences Permeability, membrane transport, intracellular transport phage Mu Porins protein export Receptors, Virus - genetics Recombinant DNA spontaneous mutants |
| title | In vivo selection and characterization of internal deletions in the lamB::lacZ gene fusion |
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