Last data update: 24 January 2024 16:39 CET
Plasmid name: pGALMFHIFNB1 (LMBP 2443)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p2443.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Saccharomyces cerevisiae α-mating factor 1 gene (MFα1); prepro secretion signal sequence (ppMF) Human interferon β cDNA (IFNB); mature sequence |
| Promoter: | Saccharomyces cerevisiae galactokinase promoter (GAL1) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pSCGAL1MF1; pSP65MFHIFNB |
| Further information: | The plasmid was constructed as follows: 1) pSCGAL1MF1 was cut with HindIII and HindII; 2) The HindIII-HindII fragment from pSP65MFHIFNB, containing the prepro secretion signal sequence (ppMF) of the Saccharomyces cerevisiae α-mating factor 1 gene (MFα1) fused to the mature human interferon β gene (hIFNB), was inserted.; 3) After HindIII digestion of the resulting intermediate construction and filling-in the sticky ends by Klenow DNA polymerase, a BamHI linker ('CCGGATCCGG') was inserted. The hIFNB gene is placed under control of the inducible GAL1 promoter. Attention: the vector part of the plasmid is probably not entirely correct: the restriction sites were not all experimentally verified. 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 at the 3' end of the GAL1 promoter is not exactly known. Other name of the plasmid is pSCGal1MFHIFNβ. |
| EMBL Accession number: | - |
| Latest sequence update: | 07/06/1994 |
| 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 Prof. Dr R. Contreras(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 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: |
pGALMFHIFNB1 (LMBP 2443) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Contreras . |
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.