Last data update: 06 May 2021 08:34 CEST
Plasmid name: pDX1 (LMBP 3375)
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|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
Baculovirus protein p35 cDNA|
|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|
|Parental clone:||pBluescriptIIKS+; pPRM-35KORF|
|Further information:||The plasmid was constructed by inserting an EcoRI fragment from pPRM-35KORF, containing full-length Baculovirus protein p35 cDNA, into the EcoRI site of pBluescriptIIKS+.
Baculovirus p35 is used as an inhibitor for apoptosis. p35 is cleaved by ICE homologs. These ICE homologs are then inhibited by the cleavage products of p35.
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 LT727473.1.
The nucleotide sequence of the Baculovirus p35 insert was provided by Dr Horvitz.
The 5' and 3' regions of the cDNA have been sequenced at the Department of Biomedical Molecular Biology (Ghent University, Belgium) with the use of M13 forward and M13 reverse primers. As a result, the sense orientation of the insert relative to the T7 promoter could also be confirmed.
|EMBL Accession number:||LT727473.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||18/10/1996|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: EcoRI and HindIII.|
|History of deposit:||This plasmid was deposited by Dr H.R. Horvitz(1) and constructed by D. Xue.
(1) Howard Hughes Medical Institute, Cambridge, USA
|Plasmid reference:||Xue et al., Nature 377 (1995), 248-251 [PMID: 7675111]
|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:||Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
pDX1 (LMBP 3375) is available at BCCM/GeneCorner. This plasmid was deposited by Dr H.R. Horvitz and constructed by D. Xue.
Howard Hughes Medical Institute, Cambridge, USA and was published in Xue et al., 1995.
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.