PurposeExpresses recombinant measles virus with duplicated P gene. wt P gene and P-F497D gene are tagged with Flag and HA peptides and si1 (siGFP) and si2 (siP) target sequences, respectively.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||58800||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Modifications to backbonePlasmid derived from p(+)MVNSe made in Roberto Cattaneo’s lab. p(+)MVNSe is derived from the original p(+)MV15894 plasmid described by Martin Billeter’s group in Radecke et al., EMBO J 14:5773 (1995) PMID:8846771. p(+)MV15894 contains the Edmonston molecular clone Genebank Z66517 with 13 point mutations including the “tag” AC>GA at positions 1818-9 of the genome, hence its acronym “Edm-tag” and a mutation in P gene resulting in 272Cyst>272Arg mutation into the V coding sequence that disable this protein. p(+)MVNSe is an elongated version of p(+)MV15894 containing the additional sequence CGTACGATGACGTCCTAG inserted just after nt 3368 to introduce unique restriction sites while preserving the genome length to compliance to the rule of six
Growth in Bacteria
Growth instructionsTransformed bacteria should never be stored at 4°C and should be kept growing at 30°C.
Copy numberHigh Copy
Gene/Insert nameMeasles virus antigenome
Insert Size (bp)17784
- Promoter T7
/ Fusion Proteins
- Flag (N terminal on insert)
- Ha (C terminal on insert)
- Cloning method Unknown
- 5′ sequencing primer AGGTGAACTccatgcGAAC
- 3′ sequencing primer TCTCTGTCATTGTGGAACTT (Common Sequencing Primers)
Unique AatII site noted in the Modifications to Backbone field was confirmed by restriction digest. Please see Notes from Addgene link to view results.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:p(+)MV-NSE-FlagrP-3g_haP-F497D-3p was a gift from Branka Horvat (Addgene plasmid # 58800 ; http://n2t.net/addgene:58800 ; RRID:Addgene_58800)