Last data update: 20 October 2020 04:14 CEST
Plasmid name: pATTrpMTNFm29S32W (LMBP 2658)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Mouse tumor necrosis factor cDNA (Tnf, TNF); L29S-R32W mutant
|Promoter:||Escherichia coli tryptophan operon promoter (trp)|
|Ribosome binding site (RBS) of the Escherichia coli tryptophan operon attenuator peptide (trp)|
|Selection marker:||Tetracycline (tet)|
|Replicon:||Escherichia coli plasmid pMB1 origin|
|Host range:||Escherichia coli; use strains expressing the trpR gene coding for the tryptophan operon repressor protein (e.g. K514).|
|Parental clone:||pATTrpMTNF; pMcTrpMTNFm29S32W|
|Further information:||This plasmid was constructed as follows : the EcoRI-PstI fragment of pMcTrpMTNFm29S32W (containing mutant MTNF)
was inserted into the unique EcoRI and PstI restriction sites of pATTrpMTNF, replacing the MTNF wild type.
The first 8 amino acids of mature MTNF are encoded by a synthetic NcoI-TaqI linker, that only differs for the first amino acid, where Leu is replaced by Val.
In the mouse TNF L29S-R32W mutant the nucleotide sequences of amino acids 29 and 32 of the wild type mature mouse TNF (CTG, coding for Leu and CGC, coding for Arg) are converted to TCG (coding for Ser) and TGG (coding for Trp).
|EMBL Accession number:||M11731, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||28/01/1993|
RBS trp | mature mouse TNF | 1 2 10 5'... GTATCGATTCC.ATG.GTA.AGA.TCA.AGT.AGT.CAA.AAT.TCG.AGT.GAC.AAG.CCT.GTA.GCC.CAC.GTC.GTA. NcoI * * ** ** TaqI Val 20 29 32 .GCA.AAC.CAC.CAA.GTG.GAG.GAG.CAG.CTG.GAG.TGG.TCG.AGC.CAG.TGG.GCC.AAC ...3' ** ** TaqI HaeIII Ser Trp
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by Dr X. Van Ostade.
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Van Ostade et al., Eur. J. Biochem. 220 (1994), 771-779 [PMID: 8143731]
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 SURE|
|Host reference:||Greener, Strategies 3 (1990), 5-6
|Cultivation medium:||LB-Lennox + tetracycline (10 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pATTrpMTNFm29S32W (LMBP 2658) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in Van Ostade et al., 1994.|
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.