| 产品名称: | pKT254omega-Ap JF311 |
|---|---|
| 商品货号: | TS158245 |
| Designations: | pKT254omega-Ap JF311 |
| Depositors: | J Frey |
| Biosafety Level: | 1 |
| Vector Information: | Size (kb): 8.8999996185302730 Vector: pKT254omega-Ap (plasmid) Construction: pHP45:omega, pKT254, pKT235 bla Marker(s):ampR,kanR Construct size (kb): 8.899999618530273 Features: marker(s): ampR, kanR replicon: ColD of KT254 terminator: T4D gene 32 |
| Applications: | integrating vector shuttle vector vector permitting positive selection for integration vector useful for interposon mutagenesis |
| Comments: | Restriction digests of the clone give the following sizes (kb): EcoRI--6.8, 2.7; HindIII--5.4, 2.6; Because resistance levels may vary, it is important to determine the proper antibiotic concentration for each strain or isolate. One of a series (ATCC 37653 through 37657) of vectors for in vitro mutagenesis (via the omega interposon) of gram-negative bacteria. An EcoRI fragment containing omega was ligated to EcoRI-digested pKT254. The spcR gene was deleted (SphI/BssHII double digest) and replaced with a 1.75 kb BamHI/BalI fragment of pKT235 containing the ampicillin resistance gene. |
| Media: | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions: | Temperature: 37.0°C |
| References: | Fellay R, et al. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of Gram-negative bacteria. Gene 52: 147-154, 1987. PubMed: 3038679 Frey J, Krisch HM. Omega mutagenesis in Gram-negative bacteria: a selectable interposon which is strongly polar in a wide range of bacterial species. Gene 36: 143-150, 1985. PubMed: 2998930 |
| Shipped: | freeze-dried |