Last data update: 24 January 2024 16:39 CET
Plasmid name: pEF-BOSbsscFvB1-2C11 (LMBP 3617)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3617.gb
(View with Genome Compiler) p3617.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse anti-BCL1 monoclonal antibody IgG1 cDNA; signal sequence (B1H) Mouse anti-BCL1 monoclonal antibody IgG1 cDNA; single chain Fv sequence (B1sFv) Hamster anti-mCD3 monoclonal antibody cDNA; single chain Fv sequence (2C11sFv) Histidine tag (His-tag); C-terminal |
| Promoter: | Human elongation factor 1α promoter (EF1α) Escherichia coli lac operon promoter Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Human granulocyte macrophage colony stimulating factor polyadenylation signal (GM-CSF polyA) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Bacteriophage M13 origin Escherichia coli plasmid pMB1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pEF-BOS; pEE12; pEE14 |
| Further information: | This plasmid allows powerful mammalian expression of the fusion protein under control of the human elongation factor 1α promoter (hEF1α). The fusion protein contains an endogenous signal sequence of the anti-BCL1 antibody (B1H) fused to the mouse IgG1 single chain Fv sequence (an antibody fragment against the antigen BCL1 (B1sFv)), and to the hamster single chain Fv sequence (an antibody fragment against the mouse CD3 (2C11sFv)). The B1sFv and 2C11sFv single chain fragments are connected by a (G4S)3 linker. After transient expression in COS-1 cells, the full-length fusion protein is secreted into the culture medium. The protein is a bispecific monovalent antibody, which can be useful for effector cell retargeting from CD3 (+) T-lymphocytes to BCL-1 (+) tumor cells. 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 LT727365.1. |
| EMBL Accession number: | LT727365.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 12/12/2001 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglII, BstEII and PstI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr J. De Jonge(1). (1) Department of Immunology-Physiology, VUB, Brussels, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Uetsuki et al., J. Biol. Chem. 264 (1989), 5791-5798 [PMID: 2564392] Mizushima et al., Nucleic Acids Res. 18 (1990), 5322 [PMID: 1698283] |
| 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] [DOI: 10.1016/s0022-2836(83)80284-8] 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) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pEF-BOSbsscFvB1-2C11 (LMBP 3617) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. De Jonge. |
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.