GREAT AT SMALL THINGS

0

LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPLT10HIs (LMBP 2589)

Add to cart

New search

Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p2589.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Escherichia coli outer membrane protein II gene (ompA); signal sequence
Human immunoglobulin γ3; heavy chain hinge region
Promoter: Phage λ major leftward promoter (λ PL)
Phage T7 gene 10 promoter (T7g10)
Ribosome
binding site:
Ribosome binding site (RBS) of the phage T7 gene 10 (T7g10)
Terminator: Phage T7 gene 10 terminator (T7g10)
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
Parental clone: pPLT10HB2Mms; pMacBLAL2HI
Further information: The plasmid was constructed by substituting the SacI (nucleotide position 167) - BstXI (blunted with T4 DNA polymerase) fragment of pPLT10HB2Mms, encoding the mutated mature human β2 microglobulin gene, by the DraI (nucleotide position 1700) - SacI fragment of pMacBLAL2HI, encoding the human γ3 hinge region. In this way, the human γ3 hinge region (HI) was fused in phase to the signal sequence of the E. coli outer membrane protein II (sompA).
The translation of the fusion protein terminates two amino acids behind the fourth hinge region (H4), caused by the presence of a termination codon in the CH2 domain.
For T7 driven expression use a strain containing a controllable T7 RNA polymerase gene, as well as a cI function: preferably a pT7POL plasmid (Mertens et al., Biotechnology 13 (1995), 175-179) or e.g. BL21(DE3)[pcI857]. Proceed as follows: first transform auxiliary plasmid, make competent cells again and then transform the expression plasmid.
The sequence of the inserted hIG3 fragment was taken from 'GENBANK Release 55; Locus: HUMIGHAF' and contains some non-identified nucleotides (indicated by N).
Other name of the plasmid is pPLT10HIsT.
EMBL Accession number: J00231, view at EMBL, GenBank, DDBJ
Latest sequence update: 23/01/1997
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr K. De Sutter(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
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)
Cultivation temperature: 28°C
Biosafety level: L1
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.

New search