Last data update: 24 November 2020 04:18 CET
Plasmid name: pPLc3 (LMBP 2470)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human Hepatitis B core protein gene (HBc); synthetic sequence|
|Promoter:||Phage λ major leftward promoter (λ PL)|
|Ribosome binding site (RBS) of the phage MS2 polymerase gene|
|Terminator:||Synthetic polyadenylation signal (polyA)|
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin|
|Host range:||Escherichia coli; use strains with a cI function, cIts for PL controlled expression|
|Further information:||- the EcoRI site at the junction between the λ PL promoter and the MS2 DNA has been destroyed by cutting and filling in; by chance, the fill-in created an SspI site (gAATATTcc).
The plasmid is identical to pPLC3-1/1, described in Nassal (1988), with the following exceptions:
- the BglII site at position 84 of the HBc coding sequence is absent (AGATCT -> AGACCT)
- the XbaI-site at position 345 of the HBc coding sequence is absent (TCTAGA -> TATAGA)
For induction, 1 to 2 hr is optimal, longer induction periods actually reduce the yield of the core protein; the yield is not overwhelming anyhow. A 5 to 10-fold higher expression system can be obtained by deleting the Arg-rich C-terminus of the protein (from codon 145 or 149 on). However, these core particles are less stable than those from the full-length protein.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727520.1.
The nucleotide sequence of the HBc synthetic sequence corresponds with the EMBL Nucleotide Sequence Database accession number M20706.1, except for the BglII and XbaI sites as described above.
|EMBL Accession number:||M20706.1, view at EMBL, GenBank, DDBJ
LT727520.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||03/12/1991|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BglII, BglII/NcoI, EcoRI, NcoI, NcoI/XbaI, SspI and XbaI.|
|History of deposit:||This plasmid was deposited by Dr M. Nassal(1).
(1) ZMBH, Heidelberg, Germany
|Related plasmid reference:||Nassal et al., Gene 66 (1988), 279-294 [PMID: 2901997]
Koriyama et al., Hypertension 66 (2015), 167-174 [PMID: 26015450] [DOI: 10.1161/HYPERTENSIONAHA.114.04534]
Broos et al., Clin. Vaccine Immunol. 15 (2008), 852-858 [PMID: 18367580] [DOI: 10.1128/CVI.00382-07]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 MC1061(λ)|
|Host reference:||Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329]
|Related host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|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.