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| 产品名称： | pSCH2002 |
|---|---|
| 商品货号： | TS202128 |
| Designations： | pSCH2002 |
| GenBank Number： | V00618 |
| Species： | Escherichia coli (Migula) Castellani and Chalmers |
| Depositors： | KJ Shaw |
| Vector： | Construct size (kb): 3.900000095367432 DESCRIPTION OF VECTOR COMPONENT: Name of vector: pBluescript KS- Intact vector size: 2.964 Type of vector: phagemid Vector end: PstI Vector end: PstI Cloning sites: SacII XmaII NotI XbaI SpeI BamHI SmaI PstI EcoRI EcoRV HindIII ClaI (SalI HincII AccI) XhoI DraII ApaI KpnI Polylinker sites: SEE COMMENTS Construction: pUC19 Host range: Escherichia coli Features (with orientation and position when available): enhancer: none insert detection: lacZ marker(s): ampR promoter: lac, T3, T7 replicon: pMB1, f1 terminator: none Cross references: |
| Insert： | DNA: Synthetic DESCRIPTION OF INSERT COMPONENT: Genome: Escherichia coli Gene symbol: aph(3)-IIa Gene name: aminoglycoside resistance Contains complete coding sequence?: U Type of DNA: unknown Insert end: PstI Insert end: PstI Insert size (kb): 0.923 Cross references: DNA Seq. Acc.: V00618 Nucleotides 1-923 of the insert correspond to nucleotides 328-1251 of V00618. Insert lengths(kb): 0.9229999780654907 Gene product: aminoglycoside resistance aph(3)-IIa |
| Insert Size (kb)： | 0.923 |
| Media： | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Biosafety Level： | 1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Shipping Information： | Distributed: freeze-dried |
| Comments： | Restriction digests of the clone give the following sizes (kb): PstI--3.0, 1.1, 1.1; EcoRI--5.2; HindIII--5.2. The clone contains two copies of the insert. The insert contains the following restriction sites (approximate kb from the 5 end): PvuII--0.06, 0.82; SphI--0.35; NcoI--0.38; SmaI--0.79. A single gel purification of the PCR generated probe is necessary since flanking regions will co-amplify with the gene specific sequence. Failure to do so often results in high backgrounds and false positives with clinical E. coli strains. A hybridization probe may be generated using the following vector specific PCR primers: modified T3 = 5-CCCCTCACTAAAGGGAACAAAAGCTG-3 and modified T7 = 5-CGCGTAATACGACTCACTATAGGGCGAA-3. The suggested PCR generated probe encodes the last 617bp of the aph(3)-IIa gene plus 290bp of the bleomycin resistance gene from Tn5. A better probe for the aph(3)-IIa gene would be the 383bp PstI/NcoI fragment. |
| Classification： | Enterobacteriaceae, Escherichia |
| References： | Shaw KJ, et alThe application of molecular techniques for the study of aminoglycoside resistanceIn: Shaw KJ, et alMethods in molecular medicine: molecular approaches for the diagnosis and investigation of bacterial diseasesTotowa, NJHumana Presssubmitted, 1996 |