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| 产品名称： | pHK9 |
|---|---|
| 商品货号： | TS145173 |
| Designations： | pHK9 |
| Depositors： | F Spencer |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 10.8999996185302700 Vector: pHK9 (plasmid) Marker(s):ampR Construct size (kb): 10.89999961853027 Features: marker(s): ampR, LYS2 replicon: ori MCS: SalI (delta15) restriction site: EcoRI,BamHI,XhoI,HpaI,NruI,BglII,PstI restriction site: PvuII coding sequence: karldelta15 |
| Applications： | cloning vector |
| Comments： | Restriction digests of the clone give the following sizes (kb): BamHI--10.9; EcoRI--6.5, 2.4, 2.05; PstI--9.0, 1.8; PvuII--10.9. Restriction digests of the clone give the following sizes (kb): BglII--7.8; EcoRI--7.8; HindIII--5.2, 2.5; EcoRI/HindIII--5.2, 2.5. Supports introduction of kar1 delta 15 mutation into lys2 yeast strains by two step gene replacement. The karl delta 15 mutation supports transfer of YAC clones between yeast strains by Karl-mating (REF). To integrate pHK9, linearize within the kar1 gene at the unique PvuII site. Cells that have lost the LYS2 marker through mitotic excision can be selected on alpha- aminoadipate. |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Vallen EA, et al. Separate domains of KAR1 mediate distinct functions in mitosis and nuclear fusion. J. Cell Biol. 117: 1277-1287, 1992. PubMed: 1607389 Spencer F, et al. Yeast kar1 mutants provide an effective method for YAC transfer to new hosts. Genomics 22: 118-126, 1994. PubMed: 7959757 Hugerat Y, et al. A versatile method for efficient YAC transfer between any two strains. Genomics 22: 108-117, 1994. PubMed: 7959756 Spencer F, Simchen GTransfer of YAC clones to new hosts by karyogamy deficient mating.In: Spencer F, Simchen GCurrent Protocols in Human GeneticsNew York, N.Y.John Wiley and Sonsin press, 1995 |
| Shipped： | freeze-dried |