Last data update: 24 November 2020 04:18 CET
Plasmid name: pPL8 (LMBP 3339)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human thymidine phosphorylase gene (hTP; human platelet derived endothelial cell growth factor)|
|Promoter:||Phage T7 gene 10 promoter (T7g10)
Phage T3 promoter
Escherichia coli lac operon promoter
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Further information:||The plasmid was constructed by inserting an approx. 1.8 kb EcoRI cDNA fragment, encoding the human platelet-derived endothelial cell growth factor/thymidine phosphorylase gene (hPDECGF or hTP).
Restriction enzyme analysis shows a BamHI fragment of approx. 1100 bp, which indicates that the gene is inserted in sense orientation relative to the T7 promoter in pBluescriptIIKS or antisense relative to the T7 promoter in pBluescriptIISK (the datasheet of Dr. Miyazono shows an insert sense to T7 in pBluescriptIISK).
The orientation of the f1ori is not known.
The first (approx.) 100 bp of the 5' untranslated region are not present in the cDNA. The length of the 3' dA tail and of the dC/dG regions at the 5' and 3' ends are not known.
The cDNA codes for a functional thymidine phosphorylase protein.
The gene can be excised as an EcoRI fragment.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727468.1.
Other name of the plasmid is pPL5.
|EMBL Accession number:||LT727468.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||21/05/1996|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: ApaI, BamHI, EcoRI and NcoI.|
|History of deposit:||This plasmid was deposited by Dr Kohei Miyazono(1) and provided under conditions of restricted use as stated by Dr Carl-Hendrik Heldin(2).
(1) The Cancer Institute, Tokyo, Japan
(2) Ludwig Institute for Cancer Research, Uppsala, Sweden
|Plasmid reference:||Ishikawa et al., Nature 338 (1989), 557-562 [PMID: 2467210]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061|
|Host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Related host reference:||Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.