Last data update: 28 October 2020 04:17 CET
Plasmid name: pCAGGS-E-hPTBdelnls (LMBP 5259)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human polypyrimidine tract binding protein 1 cDNA (PTBP1, HNRNP-I, HNRPI, pPTB, PTB-1, PTB2, PTB3, PTB4, GeneID 5725); fragment
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
|Terminator:||Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
|Host range:||Escherichia coli
Mammalian cells; SV40 permissive cells
|Further information:||The plasmid was constructed as follows: 1) the human PTBΔnls cDNA was amplified by PCR; 2) the NotI/XhoI digested PCR amplicon was fused to the E-tag of a NotI/XhoI opened pCAGGS-based expression plasmid.
pCAGGS-E-hPTBdelnls is useful for highly efficient expression of human PTBΔnls under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
The PTBΔnls fragment, lacking the nuclear localization sequence, is expressed exclusively in the cytoplasm, whereas the full-length PTBP1 protein resides in the nucleus and the cytoplasm.
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 LT726989.1.
The nucleotide sequence of the human PTBP1 cDNA corresponds with the Genbank Nucleotide Sequence Database accession number NM_002819.4.
Other name of the plasmid is pCAGGS E PTB del NLS.
|EMBL Accession number:||NM_002819.4, view at GenBank
LT726989.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||02/02/2007|
Primers used to amplify the hPTBdelnls coding sequence: forward: 5' ATAAGAAAGCGGCCGCTATGGTCCCCTCTAGAGTGATCCACATCC 3' NotI reverse: 5' AACCGCTCGAGCTAGATGGTGGACTTGGAGAAGGAGACCCG 3' XhoI
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: NcoI, NotI and NotI/XhoI.|
|History of deposit:||This plasmid was deposited by Y. Bruynooghe(1) (2) and Prof. Dr R. Beyaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Cornelis et al., Nucleic Acids Res. 33 (2005), 3095-3108 [PMID: 15928332]
|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:||-|
Refer in your Materials and Methods:
|pCAGGS-E-hPTBdelnls (LMBP 5259) is available at BCCM/GeneCorner. This plasmid was deposited by Y. Bruynooghe and Prof. Dr R. Beyaert and was published in Cornelis et al., 2005.|
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.