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| 产品名称： | pGK::nucMCS plasmid in Lactococcus lactis subsp. lactis |
|---|---|
| 商品货号： | TS142868 |
| Designations： | pGK::nucMCS plasmid in Lactococcus lactis subsp. lactis |
| Depositors： | Y Le Loir, P Langella |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 4.30 Vector: pGK::nucMCS (plasmid) Promoters: Promoter nuc Construction: pGK12, nuc Marker(s):eryR Construct size (kb): 4.30 Features: leader peptide: nuc leader marker(s): eryR promoter: nuc replicon: pWV01 MCS: NsiI...XhoI ribosome-binding site: nuc coding sequence: nuc (processed) |
| Applications： | vector permitting visual detection of recombinants |
| Comments： | To detect nuclease activity, use TBD agar: 100 ml 0.05 M Tris, pH 9; 1 g agar; 1 g NaCl; 10 ul CaCl2, 1 M; ssDNA (0.5% w/v). Heat until DNA and agar are dissolved and melted. Add 9 mg Toluidine blue O dye (Sigma T3260) and heat again. Melt for use. Replicates as a rollling circle to a level of 20-30 copies per chromosome in L. lactis. High copy-number plasmid vector for gram positive bacteria. Permits identification of recombinants by detecting secreted nuclease activity. Phenotype(s) verified: eryR |
| Media： | ATCC® Medium 1640: LB medium (ATCC medium 1065) with 25 mcg/ml erythromycin |
| Growth Conditions： | Temperature: 30.0°C |
| References： | Le Loir Y, et al. Direct screening of recombinants in gram-positive bacteria using the secreted staphylococcal nuclease as a reporter. J. Bacteriol. 176: 5135-5139, 1994. PubMed: 8051029 Lachica RV, et al. Metachromatic agar diffusion methods for detecting staphyloccal nuclease activity. Appl. Microbiol. 21: 585-587, 1971. PubMed: 4930272 Kok J, et al. Construction of plasmid cloning vectors for lactis streptococci which also replicate in Bacillus subtilis and Escherichia coli. Appl. Environ. Microbiol. 48: 726-731, 1984. PubMed: 6095756 |
| Shipped： | frozen |