pKS3 [KS334] (ATCC® 53583)

Organism: Escherichia coli (Migula) Castellani and Chalmers  /  Clone Type: Clone  /  Depositors: Harvard Univ., KL Strauch, Harvard Univ.

Permits and Restrictions

View Permits

Designations pKS3 [KS334]
Species Escherichia coli (Migula) Castellani and Chalmers
Depositors Harvard Univ., KL Strauch, Harvard Univ.
Applications
produces protein methyl-accepting chemotaxis protein I (inner membrane protein)
Vector
Construct size (kb): 6.0
DESCRIPTION OF VECTOR COMPONENT:
Name of vector: pBR322
Intact vector size: 4.363
Type of vector: plasmid
Cloning sites: EcoRI ClaI HindIII EcoRV BamHI SphI SalI XmaIII NruI BspMI
BsmI StyI AvaI BalI BspMII PvuII Tth111I NdeI AflIII PpaI
PstI PvuI ScaI SspI AatII
Polylinker sites:
Construction: pBR313
Host range: Escherichia coli
Features (with orientation and position when available):
marker(s): tetR
replicon: pMB1
marker(s): ampR
Cross references: DNA Seq. Acc.: J01749
Insert
DNA: genomic
DESCRIPTION OF INSERT COMPONENT:
Genome: Escherichia coli
Gene symbol: phoA
Gene name: alkaline phosphatase
Contains complete coding sequence?: U
Localization: 9 min
Genome: Escherichia coli
Gene symbol: tsr
Gene name: methyl-accepting chemotaxis protein I (inner membrane
protein)
Contains complete coding sequence?: U
Type of DNA: genomic
Insert end:
Insert end:
Insert size (kb):
Cross references:
Gene product: methyl-accepting chemotaxis protein I (inner membrane protein) [tsr]
Target Gene: methyl-accepting chemotaxis protein I (inner membrane protein)
Media ATCC® Medium 1896: 1065 plus ampicillin (100 mcg/ml) and XP (40 mcg/ml)
Biosafety Level 1
Shipping Information Distributed: freeze-dried
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--uncut; EcoRI--4.4, 1.55, 0.35; PstI--6.0; HindIII--6.0.
If the chimeric protein is degraded by proteases, the alkaline phosphatase becomes soluble in the periplasm and retains activity.
The host chromosome contains an insertionally inactivated gene for the periplasmic protease htrA (degP), and thus shows reduced protease activity and increased stability of periplasmic proteins.
pKS3 encodes a Tsr-alkaline phosphatase chimeric protein, with Tsr being N-terminal and anchoring the protein in the membrane. The alkaline phosphatase portion is exposed in the periplasm and retains activity.
Classification Enterobacteriaceae, Escherichia
References

Beckwith JR, Strauch KL. Periplasmic protease mutants of Escherichia coli. US Patent 4,946,783 dated Aug 7 1990

Strauch KL, Beckwith J. An Escherichia coli mutation preventing degradation of abnormal periplasmic proteins. Proc. Natl. Acad. Sci. USA 85: 1576-1580, 1988. PubMed: 3278319

Product Sheet
Product Sheet
Product Sheet