pMitoTimer
(Plasmid #52659)

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• Purpose

  Expresses mitochondrial targeted DsRed1-E5 as reporter for mitochondrial content, structure, oxidative stress and damage in vitro and in vivo

• Depositing Lab

  Zhen Yan

• Publication

  Laker et al J Biol Chem. 2014 Apr 25;289(17):12005-12015. doi: 10.1074/jbc.M113.530527( How to cite )

• Sequence Information

• Sequences (2)

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Backbone

• Vector backbone

  pDsRed2-Mito

• Backbone manufacturer

  Clontech

• Backbone size w/o insert (bp)3959

• Total vector size (bp)4658

• Modifications to backbone

  We constructed pMitoTimer by inserting the BamHI-NotI fragments of pTimer-1 (Clontech) into pDsRed2-Mito (Clontech) vector digested with BamHI and NotI using T4 ligase (New England Biolabs) following DNA isolation by using the Qiaex II gel extraction kit (Qiagen). The mitochondrial targeting sequence of the human cytochrome c oxidase subunit VIII (Cox8) gene from pDsRed2-Mito was then fused to the mutant DsRed1-E5 on the N-terminus.

• Vector type

  DsRed

• Selectable markers

  Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s)

  Kanamycin, 50 μg/mL

• Growth Temperature

  37°C

• Growth Strain(s)

  DH5alpha

• Copy number

  High Copy

Gene/Insert

• Gene/Insert name

  DsRed1-E5

• Alt name

  Timer

• Species

  Discosoma

• Insert Size (bp)

  681

• PromoterCMV

• Tag / Fusion Protein

• human cytochrome c oxidase subunit VIII mitochondrial targeting sequence (N terminal on insert)

Cloning Information

• Cloning methodRestriction Enzyme

• 5′ cloning siteBamHI (not destroyed)

• 3′ cloning siteNotI (not destroyed)

• 5′ sequencing primerCMV-F

• 3′ sequencing primerSV40pA-R (5'-GAAATTTGTGATGCTATTGC-3')

• (Common Sequencing Primers)

Resource Information

• A portion of this plasmid was derived from a plasmid made by

  Clontech

• Articles Citing this Plasmid

• 44 References

Terms and Licenses

• Academic/Nonprofit Terms

• UBMTA

• Industry Terms

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

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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.

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