Last data update: 29 October 2020 04:15 CET
Plasmid name: pCAGGSE-mIL1B-HA (LMBP 4354)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Mouse interleukin 1 beta cDNA (Il1b, Il-1b, IL-1beta, GeneID 16176)
Influenza HA epitope encoding the haemagglutinin tagging peptide; C-terminal
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
|Terminator:||Rabbit β-globin polyadenylation signal (β-globin polyA)
Simian virus 40 polyadenylation signal (SV40 polyA)
|Selection marker:||Ampicillin (amp)|
|Replicon:||Escherichia coli plasmid pMB1 origin
Simian virus 40 bidirectional origin (SV40)
|Host range:||Escherichia coli
Mammalian cells; SV40 permissive cells
|Further information:||The plasmid was constructed by PCR amplifying the mouse IL1B cds, introducing a C-terminal HA epitope, and cloning it as a NotI/XmaI fragment into the NotI/XmaI opened pCAGGS-E vector.
The plasmid is useful for highly efficient expression of the E- and HA-tagged mouse IL1B prosequence under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacIq strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The mouse IL1B insert was sequenced by BCCM/GeneCorner. The nucleotide sequence of the mouse IL1B prosequence cDNA corresponds with the Genbank accession number NM_008361.3 except for codon 185 of mouse IL1B which encodes for histidin (H) instead of tyrosin (Y), and codon 266 which consists of TCT instead of the wild type TCC.
Other names of the plasmid are pCAGGSEmproIL1βHAtag and pCAGGSEmproIL1betaHAtag.
|EMBL Accession number:||NM_008361.3, view at GenBank|
|Latest sequence update:||09/10/2013|
Primers used to amplify the HA-tagged mouse IL1B cds: <--- mIL1B ... Forward: 5' CGCAAGGAAGCGGCGGCCGC.GCA.ACT.GTT.CCT.GAA.CTC.AAC.TG NotI <--- HA-epitope ---> Reverse: 5' TCGCCCCGGG.TTA.AGC.GTA.ATC.TGG.AAC.ATC.GTA.TGG.GTA. XmaI +++ <--- mIL1B ... GGA.AGA.CAC.AGA.TTC.CAT.GGT.GAA.G +++: termination codon. Punctuation indicates reading frame.
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BamHI/SacII, BglII/EcoRI, HincII and HindIII.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(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 MC1061|
|Host reference:||Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
|Related host reference:||Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGSE-mIL1B-HA (LMBP 4354) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert .|
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.