Last data update: 19 October 2020 04:30 CEST
Plasmid name: pCAGGS-TAX1BP1-S (LMBP 3780)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human T-cell leukemia virus type I (HTLV-1) Tax1 binding protein 1 cDNA (TXBP15-1; TXBP151; TAX1BP1); small form (TAX1BP1-S)
|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
|Parental clone:||pCAGGS; pSV/TXBP15-1-AS|
|Further information:||The plasmid was constructed as follows: 1) pSV/TXBP15-1-AS was cut with EcoRI-XhoI and the (+-) 1800 bp fragment, containing the splice variant of the human TAX1BP1 cDNA, was isolated. 2) pCAGGS was cut with PvuI-EcoRI-BamHI and the (+-) 2250 bp PvuI-EcoRI fragment was isolated. 3) pCAGGS was also cut with PvuI-XhoI-SalI and the (+-) 2500 bp XhoI-PvuI fragment was isolated. 4) The three fragments were ligated, resulting in the pCAGGS-TAX1BP1-S vector.
pCAGGS-TAX1BP1-S is useful for highly efficient expression of the small human Tax1 binding protein 1 under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
The Tax1 binding protein 1 splice variant can be used in coimmunoprecipitation.
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 LT727386.1.
The TAX1BP1-S cDNA was sequenced at the Department of Biomedical Molecular Biology (Ghent University, Belgium). Referring to the sequence analysis results, the human TAX1BP1-S coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number U33821.2, except for the nucleotides CG at position 147-148 being substituted by GC, resulting in H49Q and V50L mutations, and nucleotide T at position 1203 being substituted by C, resulting in a silent mutation. These differences were confirmed by Dr K-T. Jeang (NIH, Bethesda, Maryland, USA) who deposited pSV/TXBP15-1 (LMBP 3514) and pSV/TXBP15-1-AS (LMBP 3515).
Other names of the plasmid are pCAGGSTXBP151-S and pCAGGSTAXBP151(S)s.
|EMBL Accession number:||U33821.2, view at EMBL, GenBank, DDBJ
LT727386.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||01/02/2002|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI, BglII, BglII/XbaI, EcoRI, HindII, MspA1I, PvuI/XhoI, SalI, XhoI, and XmnI.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by D. De Valck(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Related plasmid reference:||De Valck et al., Oncogene 18 (1999), 4182 - 4190 [PMID: 10435631]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH5α|
|Host reference:||Focus 8 (1986), 9
|Related host reference:||Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-TAX1BP1-S (LMBP 3780) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .|
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.