Last data update: 21 October 2021 10:13 CEST
Plasmid name: pMcMTNFm84V (LMBP 2349)
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|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
Mouse tumor necrosis factor cDNA (Tnf, TNF); A84V mutant|
|Terminator:||Phage fd terminator|
|Selection marker:||Chloramphenicol (cam)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Phage f1 origin
|Host range:||Escherichia coli|
|Further information:||The plasmid was constructed by site-specific mutagenesis of pMaMTNF, where the sequence of codon 84 (GCG) coding for Ala was replaced by GTC, coding for Val.
This mutation is described as A84V, according to the numbering of the codons in the human TNF coding sequence, although the mutated mouse codon is located on position 83.
Restriction enzyme digestions revealed that approx. 60 nucleotides are missing from the 3' untranslated region of mTNF, including the 3'-terminal (supposedly restored) EcoRI site.
After mutagenesis, use mutS strains for primary transformation (e.g. sup(-) strain WK6mutS, sup(+) strain BMH71-18mutS); for segregation of possible mutants: sup(-) strains (e.g. WK6).
The nucleotide sequence of the wild type mouse TNF corresponds with the EMBL Nucleotide Sequence Database accession number M11731.1.
|EMBL Accession number:||M11731.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||26/01/1993|
mouse TNF wild type: 80 81 82 83 84 85 86 87 5' GTC AGC CGA TTT GCT ATC TCA TAC 3' Val Ser Arg Phe Ala Ile Ser Tyr mouse TNF mutant: 80 81 82 83 84 85 86 87 5' GTC AGC CGA TTT GTG ATC TCA TAC 3' Val Ser Arg Phe Val Ile Ser Tyr
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|History of deposit:||This plasmid was deposited by Prof. Dr W. Fiers(1) (2). It was constructed by X. Van Ostade(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH1(λ)|
|Host reference:||Bachmann, in 'Escherichia coli and Salmonella: Cellular and Molecular Biology', American Society for Microbiology, Washington DC (1987), 1190-1219
|Cultivation medium:||LB-Lennox + chloramphenicol (25 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pMcMTNFm84V (LMBP 2349) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr W. Fiers .|
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.