| 产品名称： | pRE118 |
|---|---|
| 商品货号： | TS202339 |
| Designations： | pRE118 |
| Depositors： | DM Schifferli |
| Biosafety Level： | 1 |
| Vector Information： | Vector: pRE118 (plasmid)
Construction: pRE107, kanR (pUC71-K)
Size (kb): 6.20
Selectable Marker: Kanamycin resistant
Cloning sites: XbaI, KpnI, SphI, EcoRV, SacI, SmaI, EcoRI
Replicon: conditional R6K
Replicon: oriT
Marker: kanR, sacB1 (Mol Microbiol 18, 1995) |
| Applications： | Counterselection levansucrase Suicide vector Vector permitting allelic exchange of cloned inserts |
| Comments： | The sacB1 allele is a variant of sacB with certain restriction sites removed by site directed mutagenesis. Expression of sacB confers sensitivity to sucrose. The conditional R6K origin of replication requires that the pi protein be expressed in trans for plasmid maintenance. One of four allelic exchange suicide vectors (ATCC 87691 - 87694) that provide both selection for chromosomal integration (ampR, cmlR, kanR or tetR) and counterselection for loss of vector DNA and the wild type allele. Construct is suicide plasmid in any host not expressing pir. Cloned inserts may be integrated into the host chromosome (a single recombination event) following electroporation and appropriate antibiotic selection. Negative selection for sucrose sensitivity (sacB) selects for a second recombination event resulting in loss of vector DNA. |
| Media： | ATCC® Medium 1236: LB Medium (ATCC medium 1065) with 25 mcg/ml kanamycin |
| Growth Conditions： | Temperature: 37°C |
| References： | Edwards RA, et al. Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. Gene 207: 149-157, 1998. PubMed: 9511756 Kolter R, et al. Trans-complementation-dependent replication of a low molecular weight origin fragment from plasmid R6K. Cell 15: 1199-1208, 1978. PubMed: 728998 Donnenberg MS, Kaper JB. Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect. Immun. 59: 4310-4317, 1991. PubMed: 1937792 Blomfield IC, et al. Allelic exchange in Escherichia coli using the Bacillus subtilis sacB gene and a temperature-sensitive pSC101 replicon. Mol. Microbiol. 5: 1447-1457, 1991. PubMed: 1686293 McIver KS, et al. The elastase propeptide functions as an intramolecular chaperone required for elastase activity and secretion in Pseudomonas aeruginosa. Mol. Microbiol. 18: 877-889, 1995. PubMed: 8825092 Sambrook J, Russell DW. Molecular cloning: a Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2001. |
| Shipped： | frozen |
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| 产品名称： | pRE118 |
|---|---|
| 商品货号： | TS202339 |
| Designations： | pRE118 |
| Depositors： | DM Schifferli |
| Biosafety Level： | 1 |
| Vector Information： | Vector: pRE118 (plasmid)
Construction: pRE107, kanR (pUC71-K)
Size (kb): 6.20
Selectable Marker: Kanamycin resistant
Cloning sites: XbaI, KpnI, SphI, EcoRV, SacI, SmaI, EcoRI
Replicon: conditional R6K
Replicon: oriT
Marker: kanR, sacB1 (Mol Microbiol 18, 1995) |
| Applications： | Counterselection levansucrase Suicide vector Vector permitting allelic exchange of cloned inserts |
| Comments： | The sacB1 allele is a variant of sacB with certain restriction sites removed by site directed mutagenesis. Expression of sacB confers sensitivity to sucrose. The conditional R6K origin of replication requires that the pi protein be expressed in trans for plasmid maintenance. One of four allelic exchange suicide vectors (ATCC 87691 - 87694) that provide both selection for chromosomal integration (ampR, cmlR, kanR or tetR) and counterselection for loss of vector DNA and the wild type allele. Construct is suicide plasmid in any host not expressing pir. Cloned inserts may be integrated into the host chromosome (a single recombination event) following electroporation and appropriate antibiotic selection. Negative selection for sucrose sensitivity (sacB) selects for a second recombination event resulting in loss of vector DNA. |
| Media： | ATCC® Medium 1236: LB Medium (ATCC medium 1065) with 25 mcg/ml kanamycin |
| Growth Conditions： | Temperature: 37°C |
| References： | Edwards RA, et al. Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. Gene 207: 149-157, 1998. PubMed: 9511756 Kolter R, et al. Trans-complementation-dependent replication of a low molecular weight origin fragment from plasmid R6K. Cell 15: 1199-1208, 1978. PubMed: 728998 Donnenberg MS, Kaper JB. Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect. Immun. 59: 4310-4317, 1991. PubMed: 1937792 Blomfield IC, et al. Allelic exchange in Escherichia coli using the Bacillus subtilis sacB gene and a temperature-sensitive pSC101 replicon. Mol. Microbiol. 5: 1447-1457, 1991. PubMed: 1686293 McIver KS, et al. The elastase propeptide functions as an intramolecular chaperone required for elastase activity and secretion in Pseudomonas aeruginosa. Mol. Microbiol. 18: 877-889, 1995. PubMed: 8825092 Sambrook J, Russell DW. Molecular cloning: a Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 2001. |
| Shipped： | frozen |
