Last data update: 27 October 2020 04:27 CET
Plasmid name: pCAGGS-E-mMyD88s (LMBP 5646)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Mouse myeloid differentiation primary response gene 88 cDNA (Myd88, GeneID 17874); splice variant
|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 mMyd88s from the mouse macrophage cell line Mf4/4 and cloning it as a NotI/XhoI fragment into pCAGGS-E.
The plasmid contains a splice variant of mMyd88, lacking the second exon. Excision of exon 2 leads to an in-frame deletion of the complete intermediate domain (aa 110-154), resulting in a smaller protein isoform.
This plasmid is useful for highly efficient expression of mMyD88s 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 lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of the mouse Myd88 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number U84409.1.
|EMBL Accession number:||U84409.1, view at EMBL, GenBank, DDBJ|
|Latest sequence update:||18/12/2007|
Primers used to amplify the mMyD88s cds: Forward: 5' GGAATTCCCATGGGCGGCCGCG.ATG.TCT.GCG.GGA.GAC.CCC.CGC NotI *** Reverse: 5' CGCCCTGCAGCTCGAG.TCA.GGG.CAG.GGA.CAA.AGC.CTT.GGC.AAG XhoI +++ ***: start codon. +++: termination codon. Punctuation indicates reading frame.
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BstxI, NcoI/ScaI, NotI/XhoI and PvuI.|
|History of deposit:||This plasmid was deposited by Dr S. Janssens(1) (2) and 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
|Plasmid reference:||Janssens et al., Curr. Biol. 12 (2002), 467-471 [PMID: 11909531]
|Related plasmid reference:||Janssens et al., Trends Biochem. Sci. 27 (2002), 474-482 [PMID: 12217523]
Janssens et al., FEBS Lett. 548 (2003), 103-107 [PMID: 12885415]
|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:
|pCAGGS-E-mMyD88s (LMBP 5646) is available at BCCM/GeneCorner. This plasmid was deposited by Dr S. Janssens and Prof. Dr R. Beyaert and was published in Janssens et al., 2002.|
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.