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| 产品名称： | pUN20 pNN331 |
|---|---|
| 商品货号： | TS147687 |
| Designations： | pUN20 pNN331 |
| Depositors： | SJ Elledge |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 5.9000000953674320 Vector: pUN20 (plasmid) Promoters: Promoter lac Construction: pUC18 Marker(s):TRP1,ampR,SUP11 Construct size (kb): 5.900000095367432 Features: insert detection: lacZ marker(s): SUP11 marker(s): TRP1 marker(s): ampR promoter: lac replicon: ARS1 replicon: pMB1 MCS: EcoRI...HindIII centromere: CEN4 |
| Applications： | YC-type (centromeric) shuttle vector mutation detection shuttle vector vector permitting visual detection of recombinants beta-galactosidase beta-D-galactosidase |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--5.9; KpnI--4.9, 1.0; BamHI--5.9. SUP11 in the vector suppresses the red colony phenotype of ade2-101 ochre allele host strains, grown on limiting adenine media. In the absence of plasmid selection, plasmids are lost at a frequency of 1 to 3%, giving rise to red/white sectoring. One strategy for mutation scanning is to clone the normal allele of a gene into a SUP11 vector and transform into an ade2-101 host with a null mutation in the same gene, thus selecting for plasmid maintenance and producing white colonies. Vectors pUN20 (TS147687), pUN25 (ATCC 77264), pUN60 (ATCC 77266) and pUN65 (ATCC 77267) contain SUP11. A second, potentially mutated copy of the gene can then be cloned into a second centromeric shuttle vector (pUN30, ATCC 77265; pUN70, ATCC 77268; pUN90, ATCC 77269; pUN100, ATCC 77270) for transformation into the SUP11 plasmid containing host. A normal gene in plasmid #2 can relieve the selective pressure on the SUP11 plasmid, restoring the red/white sectoring; a null allele in plasmid #2 will result in SUP11 plasmid maintenance and white colonies. YC-type shuttle vector useful for the sectoring-shuffle mutagenesis assay. Also permits visual detection of recombinants by lacZalpha complementation. |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Elledge SJ, Davis RW. A family of versatile centromeric vectors designed for use in the sectoring-shuffle mutagenesis assay in Saccharomyces cerevisiae. Gene 70: 303-312, 1988. PubMed: 3063604 |
| Shipped： | freeze-dried |