TnMax — a versatile mini-transposon for the analysis of cloned genes and shuttle mutagenesis
A series of Tn7721-based mini-transposons (TnMax) has been developed which are suitable for the targeting of cloned genes, shuttle mutagenesis, identification of protein export signals and the rescue of chromosomal markers. TnMax mini-Tn consist of two 38-bp inverted repeats (IR) flanking a resoluti...
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| Veröffentlicht in: | Gene 1993-08, Vol.130 (1), p.23-31 |
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| creator | Haas, Rainer Kahrs, Andreas F. Facius, Dirk Allmeier, Helmut Schmitt, Rüdiger Meyer, Thomas F. |
| description | A series of Tn7721-based mini-transposons (TnMax) has been developed which are suitable for the targeting of cloned genes, shuttle mutagenesis, identification of protein export signals and the rescue of chromosomal markers. TnMax mini-Tn consist of two 38-bp inverted repeats (IR) flanking a resolution site (
res), a suicide replication origin (
ori
fd), and a gene conferring resistance to either chloramphenicol (TnMax1) or erythromycin (Tn
Max2). Other versions of Tn
Max (Tn
Max3and Tn
Max4) carry a promoterless alkaline phosphatase-encoding gene (
phoA') useful for the identification of protein export signals. The various mini-Tn cartridges are fused on the same plasmid with a common transposition control unit (TCU) comprising the
tnpA (transposase) and
tnpR (resolvase) genes under control of the inducible
Ptrc promoter. This configuration causes a high frequency of transposition in
Escherichia coli (approx. 10
−2 events/copy of target plasmid) and a minimum size of the mini-Tn. Like Tn
1721, Tn
Max variants prefer random insertion into plasmids rather than into the
E. colichromosome, thus representing superb tools for the insertion mutagenesis of cloned genes. The Tn
Max-borne
ori
fd simplifies the identification of targeted plasmids and facilitates shuttle mutagenesis i.e., suicide delivery of a mutated gene with subsequent allelic replacement of a corresponding resident gene, in a variety of microorganisms. Rescue of such targeted chromosomal genes is easily accomplished by the excision of Tn
Max plus flanking segments using appropriate restriction endonucleases, ligation, and transformation of an
E. colihost permissive for
ori
fd
-directed replication. |
| doi_str_mv | 10.1016/0378-1119(93)90342-Z |
| format | Article |
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res), a suicide replication origin (
ori
fd), and a gene conferring resistance to either chloramphenicol (TnMax1) or erythromycin (Tn
Max2). Other versions of Tn
Max (Tn
Max3and Tn
Max4) carry a promoterless alkaline phosphatase-encoding gene (
phoA') useful for the identification of protein export signals. The various mini-Tn cartridges are fused on the same plasmid with a common transposition control unit (TCU) comprising the
tnpA (transposase) and
tnpR (resolvase) genes under control of the inducible
Ptrc promoter. This configuration causes a high frequency of transposition in
Escherichia coli (approx. 10
−2 events/copy of target plasmid) and a minimum size of the mini-Tn. Like Tn
1721, Tn
Max variants prefer random insertion into plasmids rather than into the
E. colichromosome, thus representing superb tools for the insertion mutagenesis of cloned genes. The Tn
Max-borne
ori
fd simplifies the identification of targeted plasmids and facilitates shuttle mutagenesis i.e., suicide delivery of a mutated gene with subsequent allelic replacement of a corresponding resident gene, in a variety of microorganisms. Rescue of such targeted chromosomal genes is easily accomplished by the excision of Tn
Max plus flanking segments using appropriate restriction endonucleases, ligation, and transformation of an
E. colihost permissive for
ori
fd
-directed replication.</description><identifier>ISSN: 0378-1119</identifier><identifier>EISSN: 1879-0038</identifier><identifier>DOI: 10.1016/0378-1119(93)90342-Z</identifier><identifier>PMID: 8393825</identifier><identifier>CODEN: GENED6</identifier><language>eng</language><publisher>Lausanne: Elsevier B.V</publisher><subject>Base Sequence ; Biological and medical sciences ; Biotechnology ; catGC ; Cloning, Molecular ; DNA Mutational Analysis - methods ; DNA Repair ; DNA Transposable Elements - genetics ; DNA, Bacterial - analysis ; Escherichia coli - genetics ; Fundamental and applied biological sciences. Psychology ; Genes, Bacterial ; Genetic engineering ; Genetic technics ; Genetic Vectors ; Methods. Procedures. Technologies ; Molecular Sequence Data ; Mutagenesis, Insertional - methods ; natural transformation ; Neisseria gonorrhoeae - genetics ; Oligonucleotides ; orifd ; Promoter Regions, Genetic ; Ptrcpromoter ; shuttle-transposon ; Site specific mutagenesis ; Tn phoA ; Tn1721</subject><ispartof>Gene, 1993-08, Vol.130 (1), p.23-31</ispartof><rights>1993</rights><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-ce1edb759d8eb11e4a67ab357dec88eca8c7960610ca38b49320f3ca82aee0c13</citedby><cites>FETCH-LOGICAL-c417t-ce1edb759d8eb11e4a67ab357dec88eca8c7960610ca38b49320f3ca82aee0c13</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(93)90342-Z$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4853156$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8393825$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Haas, Rainer</creatorcontrib><creatorcontrib>Kahrs, Andreas F.</creatorcontrib><creatorcontrib>Facius, Dirk</creatorcontrib><creatorcontrib>Allmeier, Helmut</creatorcontrib><creatorcontrib>Schmitt, Rüdiger</creatorcontrib><creatorcontrib>Meyer, Thomas F.</creatorcontrib><title>TnMax — a versatile mini-transposon for the analysis of cloned genes and shuttle mutagenesis</title><title>Gene</title><addtitle>Gene</addtitle><description>A series of Tn7721-based mini-transposons (TnMax) has been developed which are suitable for the targeting of cloned genes, shuttle mutagenesis, identification of protein export signals and the rescue of chromosomal markers. TnMax mini-Tn consist of two 38-bp inverted repeats (IR) flanking a resolution site (
res), a suicide replication origin (
ori
fd), and a gene conferring resistance to either chloramphenicol (TnMax1) or erythromycin (Tn
Max2). Other versions of Tn
Max (Tn
Max3and Tn
Max4) carry a promoterless alkaline phosphatase-encoding gene (
phoA') useful for the identification of protein export signals. The various mini-Tn cartridges are fused on the same plasmid with a common transposition control unit (TCU) comprising the
tnpA (transposase) and
tnpR (resolvase) genes under control of the inducible
Ptrc promoter. This configuration causes a high frequency of transposition in
Escherichia coli (approx. 10
−2 events/copy of target plasmid) and a minimum size of the mini-Tn. Like Tn
1721, Tn
Max variants prefer random insertion into plasmids rather than into the
E. colichromosome, thus representing superb tools for the insertion mutagenesis of cloned genes. The Tn
Max-borne
ori
fd simplifies the identification of targeted plasmids and facilitates shuttle mutagenesis i.e., suicide delivery of a mutated gene with subsequent allelic replacement of a corresponding resident gene, in a variety of microorganisms. Rescue of such targeted chromosomal genes is easily accomplished by the excision of Tn
Max plus flanking segments using appropriate restriction endonucleases, ligation, and transformation of an
E. colihost permissive for
ori
fd
-directed replication.</description><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>catGC</subject><subject>Cloning, Molecular</subject><subject>DNA Mutational Analysis - methods</subject><subject>DNA Repair</subject><subject>DNA Transposable Elements - genetics</subject><subject>DNA, Bacterial - analysis</subject><subject>Escherichia coli - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes, Bacterial</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>Genetic Vectors</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Insertional - methods</subject><subject>natural transformation</subject><subject>Neisseria gonorrhoeae - genetics</subject><subject>Oligonucleotides</subject><subject>orifd</subject><subject>Promoter Regions, Genetic</subject><subject>Ptrcpromoter</subject><subject>shuttle-transposon</subject><subject>Site specific mutagenesis</subject><subject>Tn phoA</subject><subject>Tn1721</subject><issn>0378-1119</issn><issn>1879-0038</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMtuFDEQRS1EFIbAH4DkBUKwaLDb_bA3kVDEI1IQm7DJAqvaXU2MeuzB5Y7ILh_BF_IleDKjWcYbS3XPLZUOYy-keCeF7N4L1etKSmneGPXWCNXU1dUjtpK6N5UQSj9mqwPyhD0l-iXKa9v6mB1rZZSu2xX7cRm-wh_-7-4vB36DiSD7GfnaB1_lBIE2kWLgU0w8XyOHAPMteeJx4m6OAUf-EwNSCUZO10vO2_KS4X7q6Rk7mmAmfL7_T9j3Tx8vz75UF98-n599uKhcI_tcOZQ4Dn1rRo2DlNhA18Og2n5EpzU60K43neikcKD00BhVi0mVcQ2Iwkl1wl7v9m5S_L0gZbv25HCeIWBcyMqu06pvmwI2O9ClSJRwspvk15BurRR2q9VundmtM2uUvddqr0rt5X7_MqxxPJT2Hkv-ap8DOZinYs55OmCNbpVsu4Kd7jAsLm48JkvOY3A4-oQu2zH6h-_4D0G3lhk</recordid><startdate>19930816</startdate><enddate>19930816</enddate><creator>Haas, Rainer</creator><creator>Kahrs, Andreas F.</creator><creator>Facius, Dirk</creator><creator>Allmeier, Helmut</creator><creator>Schmitt, Rüdiger</creator><creator>Meyer, Thomas F.</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>7TM</scope></search><sort><creationdate>19930816</creationdate><title>TnMax — a versatile mini-transposon for the analysis of cloned genes and shuttle mutagenesis</title><author>Haas, Rainer ; Kahrs, Andreas F. ; Facius, Dirk ; Allmeier, Helmut ; Schmitt, Rüdiger ; Meyer, Thomas F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-ce1edb759d8eb11e4a67ab357dec88eca8c7960610ca38b49320f3ca82aee0c13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>catGC</topic><topic>Cloning, Molecular</topic><topic>DNA Mutational Analysis - methods</topic><topic>DNA Repair</topic><topic>DNA Transposable Elements - genetics</topic><topic>DNA, Bacterial - analysis</topic><topic>Escherichia coli - genetics</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes, Bacterial</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>Genetic Vectors</topic><topic>Methods. Procedures. Technologies</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Insertional - methods</topic><topic>natural transformation</topic><topic>Neisseria gonorrhoeae - genetics</topic><topic>Oligonucleotides</topic><topic>orifd</topic><topic>Promoter Regions, Genetic</topic><topic>Ptrcpromoter</topic><topic>shuttle-transposon</topic><topic>Site specific mutagenesis</topic><topic>Tn phoA</topic><topic>Tn1721</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Haas, Rainer</creatorcontrib><creatorcontrib>Kahrs, Andreas F.</creatorcontrib><creatorcontrib>Facius, Dirk</creatorcontrib><creatorcontrib>Allmeier, Helmut</creatorcontrib><creatorcontrib>Schmitt, Rüdiger</creatorcontrib><creatorcontrib>Meyer, Thomas F.</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>Nucleic Acids Abstracts</collection><jtitle>Gene</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Haas, Rainer</au><au>Kahrs, Andreas F.</au><au>Facius, Dirk</au><au>Allmeier, Helmut</au><au>Schmitt, Rüdiger</au><au>Meyer, Thomas F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>TnMax — a versatile mini-transposon for the analysis of cloned genes and shuttle mutagenesis</atitle><jtitle>Gene</jtitle><addtitle>Gene</addtitle><date>1993-08-16</date><risdate>1993</risdate><volume>130</volume><issue>1</issue><spage>23</spage><epage>31</epage><pages>23-31</pages><issn>0378-1119</issn><eissn>1879-0038</eissn><coden>GENED6</coden><abstract>A series of Tn7721-based mini-transposons (TnMax) has been developed which are suitable for the targeting of cloned genes, shuttle mutagenesis, identification of protein export signals and the rescue of chromosomal markers. TnMax mini-Tn consist of two 38-bp inverted repeats (IR) flanking a resolution site (
res), a suicide replication origin (
ori
fd), and a gene conferring resistance to either chloramphenicol (TnMax1) or erythromycin (Tn
Max2). Other versions of Tn
Max (Tn
Max3and Tn
Max4) carry a promoterless alkaline phosphatase-encoding gene (
phoA') useful for the identification of protein export signals. The various mini-Tn cartridges are fused on the same plasmid with a common transposition control unit (TCU) comprising the
tnpA (transposase) and
tnpR (resolvase) genes under control of the inducible
Ptrc promoter. This configuration causes a high frequency of transposition in
Escherichia coli (approx. 10
−2 events/copy of target plasmid) and a minimum size of the mini-Tn. Like Tn
1721, Tn
Max variants prefer random insertion into plasmids rather than into the
E. colichromosome, thus representing superb tools for the insertion mutagenesis of cloned genes. The Tn
Max-borne
ori
fd simplifies the identification of targeted plasmids and facilitates shuttle mutagenesis i.e., suicide delivery of a mutated gene with subsequent allelic replacement of a corresponding resident gene, in a variety of microorganisms. Rescue of such targeted chromosomal genes is easily accomplished by the excision of Tn
Max plus flanking segments using appropriate restriction endonucleases, ligation, and transformation of an
E. colihost permissive for
ori
fd
-directed replication.</abstract><cop>Lausanne</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>8393825</pmid><doi>10.1016/0378-1119(93)90342-Z</doi><tpages>9</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0378-1119 |
| ispartof | Gene, 1993-08, Vol.130 (1), p.23-31 |
| issn | 0378-1119 1879-0038 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_16683754 |
| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Base Sequence Biological and medical sciences Biotechnology catGC Cloning, Molecular DNA Mutational Analysis - methods DNA Repair DNA Transposable Elements - genetics DNA, Bacterial - analysis Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Genes, Bacterial Genetic engineering Genetic technics Genetic Vectors Methods. Procedures. Technologies Molecular Sequence Data Mutagenesis, Insertional - methods natural transformation Neisseria gonorrhoeae - genetics Oligonucleotides orifd Promoter Regions, Genetic Ptrcpromoter shuttle-transposon Site specific mutagenesis Tn phoA Tn1721 |
| title | TnMax — a versatile mini-transposon for the analysis of cloned genes and shuttle mutagenesis |
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