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pGK::nucMCS plasmid in Lactococcus lactis subsp. lactis
pGK::nucMCS plasmid in Lactococcus lactis subsp. lactis
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編號:B234062
品牌:Mingzhoubio

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RNA

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產(chǎn)品名稱
pGK::nucMCS plasmid in Lactococcus lactis subsp. lactis
商品貨號
B234062
Designations
pGK::nucMCS plasmid in Lactococcus lactis subsp. lactis
Depositors
Y Le Loir, P Langella
培養(yǎng)基
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

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