Last data update: 20 October 2020 04:14 CEST
Plasmid name: pBMGNeo (LMBP 1971)
|Price category:||Cat. 3 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Promoter:||Mouse metallothionein 1 promoter (Mt1)
Herpes simplex virus (HSV) thymidine kinase promoter (TK)
|Terminator:||Herpes simplex virus (HSV) thymidine kinase polyadenylation signal (TK polyA)
Rabbit β-globin polyadenylation signal (β-globin polyA)
|Selection marker:||Ampicillin (amp)
Neomycin (neo; G418; kanamycin (kan))
|Replicon:||Escherichia coli plasmid pMB1 origin
Bovine papilloma virus origin (BPV)
|Host range:||Escherichia coli
Mammalian cells; e.g. NIH3T3, FR3T3, C127
|Further information:||pBMGNeo has the ability to replicate extrachromosomally in some mammalian cells (because of the 69% fragment of bovine papilloma virus) as well as in prokaryotic cells (due to the E. coli pMB1 origin of replication).
The bacterial gene for neomycin resistance is used as a dominant selectable marker: in bacteria, the expression of this gene is driven by its own promoter; in mammalian cells by the HSV-TK promoter.
The mouse Mt1 promoter controls the expression of inserted cDNA's in mammalian cells; this mouse Mt1 promoter can be induced by some kations such as Zn(2)(+) or Cd(2)(+).
The polyA signal and the 3' flanking region from the rabbit β-globin gene allow efficient processing and termination of transcription.
The fragment of the human genomic β-globin DNA carries an activity that enhances the ability of BPV vectors to replicate as episomes.
cDNA can be inserted into the SalI expression site as well as in the XhoI expression site. The majority of full-length cDNA's can be inserted since XhoI and SalI sites are rare in mammalian DNA.
Because of the presence of the second intron of the rabbit β-globin gene, cDNA inserts in the SalI site (located at 20 bp in front of the splice donor signal) are spliced in the 3' untranslated region and inserts in the XhoI site (located at 50 bp behind the splice acceptor signal) in the 5' untranslated region.
The nucleotide sequence of the plasmid was compiled based on the information in Karasuyama et al. (1988). However, we could not completely trace back the sequence, that thus contains some uncertainties. Consequently, there is also uncertainty as to the cutting frequency of the restriction enzymes, except for the sites that were analysed in the authenticity test.
The nucleotide sequence of the bovine papilloma virus corresponds with the EMBL Nucleotide Sequence Database accession number X02346.1.
Name mentioned in Karasuyama et al. (1988) is BMGNeo.
|EMBL Accession number:||X02346.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||09/09/2008|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI, HindII/HindIII, NotI, NotI/XbaI, SalI, XbaI and XhoI.
This plasmid has also been fully sequenced but the NGS sequence data still need to be implemented.
|History of deposit:||This plasmid was deposited by Prof. Dr F. Melchers(1).
(1) Basel Institute for Immunology, Switserland
|Plasmid reference:||Karasuyama et al., Eur. J. Immunol. 18 (1988), 97-104 [PMID: 2831066]
|Related plasmid reference:||Meneguzzi et al., EMBO J. 3 (1984), 365-371 [PMID: 6325168]
Sambrook et al., EMBO J. 4 (1985), 91-103 [PMID: 2990897]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12xB HB101|
|Host reference:||Boyer and Roulland-Dussoix, J. Mol. Biol. 41 (1969), 459-472 [PMID: 4896022]
|Related host reference:||Sambrook et al. (eds), Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, NY (1989) [ISSN/ISBN: 0879693096]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pBMGNeo (LMBP 1971) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr F. Melchers and was published in Karasuyama et al., 1988.|
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.