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| 产品名称： | pdeltaADE2 |
|---|---|
| 商品货号： | TS151111 |
| Designations： | pdeltaADE2 |
| Depositors： | JD Boeke |
| Biosafety Level： | 1 |
| Host： | Escherichia coli MC1066 |
| Vector Information： | Size (kb): 8.4 DESCRIPTION OF VECTOR: Intact vector size: 8.400 Type of vector: plasmid Cloning sites: Polylinker sites: Construction: pBluescript, URA3, hisG, ADE2 sequences Host range: Saccharomyces cerevisiae; Escherichia coli Features (with orientation and position when available): restriction site: BamHI other: ADE2 flanking sequence coding sequence: hisG, -> marker(s): URA3, -> coding sequence: hisG, -> other: ADE2 flanking sequence restriction site: BamHI replicon: pMB1 marker(s): ampR Vector: pdeltaADE2 (plasmid) Construction: pBluescript, URA3, hisG, ADE2 sequences Marker(s):URA3,ampR Construct size (kb): 8.4 Features: marker(s): URA3 marker(s): ampR other: ADE2 flanking sequence replicon: pMB1 restriction site: BamHI coding sequence: hisG |
| Applications： | contains sequence ATP phosphoribosyltransferase marker deletion vector phosphoribosylaminoimidazole succinocarboxamide synthetase SAICAR synthetase, phosphoribosylaminoimidazole carboxylase produces protein uridine monophosphate synthetase UMP synthase, orotate phosphoribosyltransferase, orotidine 5-phosphate decarboxylase, orotate phosphoribosyltransferase 1 |
| Comments： | Restriction digests of the clone give the following sizes (kb): BamHI--5.2, 3.2; EcoRI--5.0, 3.4; HindIII--7.0, 1.3. The two step selection process requires a ura3 transformation host (this host can be created using pJL164 (ATCC 87471)). After transformation with the BamHI digested plasmid, URA3 integrants are selected on ura- plates. The designer deletion strain is then recovered by selection on 5-FOA plates (loss of URA3 and ADE2 markers by a homologous recombination event between the two hisG repeats). The deleted host retains the coding sequence for six C-terminal amino acids of ADE2. E. coli containing this plasmid should be grown on media lacking pyrimidines to select for URA3-containing cells. This deleter vector is used to create designer yeast strains with a non-revertable ade2 auxotrophic marker deletion. The 5.2 kb BamHI insert contains two direct repeats of the Salmonella hisG gene flanking URA3 and about 700 bp of homology to sequences upstream and downstream of the ADE2 gene flanking the hisG-URA3-hisG sequence. |
| Media： | ATCC® Medium 2057: M9 salts with supplements |
| Growth Conditions： | Temperature: 37°C |
| References： | Aparicio OM, et al. Modifiers of position effect are shared between telomeric and silent mating-type loci in S. cerevisiae. Cell 66: 1279-1287, 1991. PubMed: 1913809 Alani E, et al. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics 116: 541-545, 1987. PubMed: 3305158 Jef D Boeke, personal communication |
| Related Products： | component of:ATCC 87472 |
| Shipped： | frozen |
| Shipping Information： | Distributed: freeze-dried |