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URL: http://www.addgene.org/26809

Proper Citation: RRID:Addgene_26809

Insert Name: MyoD-ER(T)

Organism: Mus musculus

Bacterial Resistance: Ampicillin

Defining Citation: PMID:18511457

Vector Backbone Description: Backbone Size:6707; Vector Backbone:pRRL-cPPT-X-PRE-SIN; Vector Types:Lentiviral; Bacterial Resistance:Ampicillin

Comments: Cloning of the MyoD-ER(T): Briefly, the full-length mouse MyoD cDNA was isolated and cloned from a previously described adenoviral vector (Lattanzi et al., 1998). A cDNA for the modified estrogen receptor responsive to tamoxifen and 4-hydroxytamoxifen, ER(T) (Danielian et al., 1993), was cloned using a PCR method with the primer pair including an internal ClaI restriction enzyme site: (sense) 5′-ACGGCATCGATTCTGCTGGAGACATGAGAGCT-3′, and (anti-sense) 5′-GCGCCATCGATGACTGTGGCAGGGAAACCCT-3′, and then inserted into the NarI site (amino acid 173) in the middle of the mouse MyoD cDNA. The final clone was verified by DNA sequencing. There is a mutation G525R in the mouse estrogen receptor; the mutation confers selective 4-hydroxytamoxifen sensitivity (vs. estradiol) (Danielian et al. 1993 Molec Endocrin 7: 232-240).

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