 | | | | pGPS4 Transprimer Donor | | | Will be discontinued as of April 1, 2009 | |
| |
|
|
 |
|
Prices are in US dollars and valid only for US orders.
|
Description:
pGPS4 is the Transprimer-4 donor plasmid used in the GPS™-LS Linker Scanning System (NEB #E7102S). TnsABC* Transposase removes the Transprimer-4 element from pGPS4 and inserts it randomly into the target DNA in vitro (1-3). Digestion with PmeI removes the majority of Transprimer-4. Religation results in a 15 bp insertion which includes a PmeI site. Transprimer-4 encodes chloramphenicol-resistance. This double-stranded plasmid is 3899 bp in size.
Source:
pGPS4 is isolated from E. coli BW23474, an E. coli strain that expresses the pir-116 replication factor (4), by a standard plasmid purification procedure.
Concentration:
20 μg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 8.0 @ 25°C
Storage Temperature:
-20°C
Notes
General notes:- pGPS4 will not replicate in ordinary lab strains of E. coli. Its origin of replication is from the transmissible plasmid R6K, and it depends on a replication initiation protein of plasmid origin (the protein product of the pir gene) not normally present in laboratory strains (4,5).
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
GPS Control Reaction::
A 20 µl GPS Control Reaction was performed using 0.02 µg pGPS4,
0.08 µg Contol Target plasmid and 1 µl TnsABC* Transposase in 1X GPS Buffer supplemented with 15 mM Start Solution after 10 minutes. The reaction was incubated for 1 hour at 37°C and transformed into chemically-competent E. coli (1 x 107 per µg pBR322). Less than 1% of the resulting chloramphenicol-resistant colonies contained cointegrate molecules.
References
- Craig, N.L. (1996) Curr. Top. Microbiol. Immunol., 204, 27-48.
- Stellwagen, A.E. and Craig, N.L. (1997) Genetics, 145, 573-585.
- Biery, M.C., Stewart, F.J., Stellwagen, A.E., Raleigh, E.A. and Craig, N.L. (2000) Nucl. Acids Res., 28, 1067-1077.
- Metcalf, W.W., Jiang, W. and Wanner, B.L. (1994) Gene, 138, 1-7.
- Kolter, R., Inuzuka, M. and Helinski, D.R. (1978) Cell, 15, 1199-1208.
Companion Products
GPS™ Buffer Pack
GPS™-1 Genome Priming System
GPS™-LS Linker Scanning System
GPS™-M Mutagenesis System
pGPS1.1 Transprimer Donor
pGPS2.1 Transprimer Donor
pGPS3 Transprimer Donor
pGPS5 Transprimer Donor
TnsABC* Transposase
Legal
Patents:
Johns Hopkins University: Licensed Under U.S. Patent No. 6,420,524
|
