| 产品名称: | pKB803 |
|---|---|
| 商品货号: | TS159488 |
| Designations: | pKB803 |
| Depositors: | OmniGene, Inc., KC Backman, OmniGene, Inc. |
| U.S. Patent: | |
| Disclosure: | This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished. This material may not have been produced or characterized by ATCC. |
| Biosafety Level: | 1 |
| Host: | Distribution host: Escherichia coli YMC9 |
| Vector Information: | Size (kb): 9.3000001907348630 DESCRIPTION OF VECTOR COMPONENT: Name of vector: pKB800 Intact vector size: 8.100 Type of vector: plasmid Vector end: XhoI Vector end: XhoI Cloning sites: XhoI Polylinker sites: Construction: pBR322, pKB720 Host range: Escherichia coli Features (with orientation and position when available): restriction site: KpnI repressor gene: cI857 restriction site: XhoI coding sequence: N coding sequence: xis coding sequence: int operator: att restriction site: KpnI replicon: pMB1 marker(s): ampR Cross references: Vector: pKB803 (plasmid) Construction: pKB800, tyrA Marker(s):ampR Construct size (kb): 9.300000190734863 Features: marker(s): ampR operator: att replicon: pMB1 repressor gene: cI857 restriction site: KpnI restriction site: XhoI coding sequence: N coding sequence: int coding sequence: tyrA coding sequence: xis |
| Applications: | host modification produces protein chorismate mutase T produces protein prephenate dehydrogenase |
| Comments: | Restriction digests of the clone give the following sizes (kb): KpnI--7.2, 2.1; XhoI--8.2, 1.2; ClaI--9.0, 0.4. Integration can be achieved by deletion of the KpnI fragment of the vector (containing the origin of replication and ampicillin resistance gene), followed by transformation of a tyrA deficient host and selection for tyrosine prototrophy. A host lacking pheA-tyrA-aroF is recommended. Integration efficiency can be improved by co-infection of the transformed cells with an integration-deficient helper phage, such as lambda(imm434b104). After temperature induction at 42 C, the tyrA gene will be excised from the chromosome and lost, effectively stopping cell growth in tyrosine deficient media. Vector allowing insertion of the tyrA gene into the E. coli chromosome and controlled excision of the same DNA upon temperature induction. |
| Media: | ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin |
| Growth Conditions: | Temperature: 30.0°C |
| References: | Backman KC. Controlled gene excision. US Patent 4,743,546 dated May 10 1988 |
| Shipped: | freeze-dried |