A plasmid vector for isolation of strong promoters in Escherichia coli
In order to isolate very strong promoters from bacteria and bacteriophage a plasmid named pProm was constructed. It possesses an origin (ORI) for replication in Gram-negative bacteria, an ORI for replication in Gram-positive bacteria, a promoterless ampicillin resistance gene with a multiple cloning...
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| Formato: | Artículo publishedVersion |
| Lenguaje: | Inglés |
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1999
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_03781097_v177_n1_p75_Carbonelli |
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paperaa:paper_03781097_v177_n1_p75_Carbonelli2023-06-12T16:47:59Z A plasmid vector for isolation of strong promoters in Escherichia coli FEMS Microbiol. Lett. 1999;177(1):75-82 Carbonelli, D.L. Corley, E. Seigelchifer, M. Zorzópulos, J. Bacterial promoter Bacteriophage promoter Promoter isolation Strong promoter Vector ampicillin DNA fragment penicillinase plasmid vector tetracycline antibiotic resistance article bacteriophage DNA sequence Escherichia coli nonhuman nucleotide sequence penicillin resistance plasmid priority journal promoter region Staphylococcus aureus Bacteria (microorganisms) bacteriophage a Escherichia coli Escherichia coli Negibacteria Posibacteria Prokaryota Staphylococcus aureus Staphylococcus aureus unidentified bacteriophage In order to isolate very strong promoters from bacteria and bacteriophage a plasmid named pProm was constructed. It possesses an origin (ORI) for replication in Gram-negative bacteria, an ORI for replication in Gram-positive bacteria, a promoterless ampicillin resistance gene with a multiple cloning site (MCS) in the position formerly occupied by the ampicillin promoter, a tetracycline resistance gene for selection in Gram-negative bacteria and a chloramphenicol resistance gene for selection in Gram-positive bacteria. Insertion in the MCS of DNA fragments of Staphylococcus aureus bacteriophages resulted in isolation of several clones very resistant to ampicillin. The DNA fragments inserted in these recombinant plasmids were sequenced and all of them contained putative promoter motifs. Direct measurement of the penicillinase activity indicated that one of the isolated promoters could be included within a group of the stronger known prokaryotic promoters. According to these results pProm is a powerful tool to perform studies on promoter strength and for industrial applications. Copyright (C) 1999 Federation of European Microbiological Societies. Fil:Seigelchifer, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1999 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_03781097_v177_n1_p75_Carbonelli |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| language |
Inglés |
| orig_language_str_mv |
eng |
| topic |
Bacterial promoter Bacteriophage promoter Promoter isolation Strong promoter Vector ampicillin DNA fragment penicillinase plasmid vector tetracycline antibiotic resistance article bacteriophage DNA sequence Escherichia coli nonhuman nucleotide sequence penicillin resistance plasmid priority journal promoter region Staphylococcus aureus Bacteria (microorganisms) bacteriophage a Escherichia coli Escherichia coli Negibacteria Posibacteria Prokaryota Staphylococcus aureus Staphylococcus aureus unidentified bacteriophage |
| spellingShingle |
Bacterial promoter Bacteriophage promoter Promoter isolation Strong promoter Vector ampicillin DNA fragment penicillinase plasmid vector tetracycline antibiotic resistance article bacteriophage DNA sequence Escherichia coli nonhuman nucleotide sequence penicillin resistance plasmid priority journal promoter region Staphylococcus aureus Bacteria (microorganisms) bacteriophage a Escherichia coli Escherichia coli Negibacteria Posibacteria Prokaryota Staphylococcus aureus Staphylococcus aureus unidentified bacteriophage Carbonelli, D.L. Corley, E. Seigelchifer, M. Zorzópulos, J. A plasmid vector for isolation of strong promoters in Escherichia coli |
| topic_facet |
Bacterial promoter Bacteriophage promoter Promoter isolation Strong promoter Vector ampicillin DNA fragment penicillinase plasmid vector tetracycline antibiotic resistance article bacteriophage DNA sequence Escherichia coli nonhuman nucleotide sequence penicillin resistance plasmid priority journal promoter region Staphylococcus aureus Bacteria (microorganisms) bacteriophage a Escherichia coli Escherichia coli Negibacteria Posibacteria Prokaryota Staphylococcus aureus Staphylococcus aureus unidentified bacteriophage |
| description |
In order to isolate very strong promoters from bacteria and bacteriophage a plasmid named pProm was constructed. It possesses an origin (ORI) for replication in Gram-negative bacteria, an ORI for replication in Gram-positive bacteria, a promoterless ampicillin resistance gene with a multiple cloning site (MCS) in the position formerly occupied by the ampicillin promoter, a tetracycline resistance gene for selection in Gram-negative bacteria and a chloramphenicol resistance gene for selection in Gram-positive bacteria. Insertion in the MCS of DNA fragments of Staphylococcus aureus bacteriophages resulted in isolation of several clones very resistant to ampicillin. The DNA fragments inserted in these recombinant plasmids were sequenced and all of them contained putative promoter motifs. Direct measurement of the penicillinase activity indicated that one of the isolated promoters could be included within a group of the stronger known prokaryotic promoters. According to these results pProm is a powerful tool to perform studies on promoter strength and for industrial applications. Copyright (C) 1999 Federation of European Microbiological Societies. |
| format |
Artículo Artículo publishedVersion |
| author |
Carbonelli, D.L. Corley, E. Seigelchifer, M. Zorzópulos, J. |
| author_facet |
Carbonelli, D.L. Corley, E. Seigelchifer, M. Zorzópulos, J. |
| author_sort |
Carbonelli, D.L. |
| title |
A plasmid vector for isolation of strong promoters in Escherichia coli |
| title_short |
A plasmid vector for isolation of strong promoters in Escherichia coli |
| title_full |
A plasmid vector for isolation of strong promoters in Escherichia coli |
| title_fullStr |
A plasmid vector for isolation of strong promoters in Escherichia coli |
| title_full_unstemmed |
A plasmid vector for isolation of strong promoters in Escherichia coli |
| title_sort |
plasmid vector for isolation of strong promoters in escherichia coli |
| publishDate |
1999 |
| url |
http://hdl.handle.net/20.500.12110/paper_03781097_v177_n1_p75_Carbonelli |
| work_keys_str_mv |
AT carbonellidl aplasmidvectorforisolationofstrongpromotersinescherichiacoli AT corleye aplasmidvectorforisolationofstrongpromotersinescherichiacoli AT seigelchiferm aplasmidvectorforisolationofstrongpromotersinescherichiacoli AT zorzopulosj aplasmidvectorforisolationofstrongpromotersinescherichiacoli AT carbonellidl plasmidvectorforisolationofstrongpromotersinescherichiacoli AT corleye plasmidvectorforisolationofstrongpromotersinescherichiacoli AT seigelchiferm plasmidvectorforisolationofstrongpromotersinescherichiacoli AT zorzopulosj plasmidvectorforisolationofstrongpromotersinescherichiacoli |
| _version_ |
1769810184250589184 |