PCx35IL-pET42
(Plasmid #42916)

• Depositing Lab: John O'Brien
• Publication: Burr et al Biochem Biophys Res Commun. 2005 Oct 7;335(4):1191-8.

Backbone

• Vector backbone: pET42a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5930
• Total vector size (bp): 6133
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Requires a DE3 strain for expression of protein.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Connexin 35 cytoplasmic loop
• Alt name: Cx35
• Alt name: Cx36
• Alt name: gjd2
• Species: Perch (Morone americana)
• Insert Size (bp): 240
• Mutation: cytoplasmic loop is aa 101-179
• GenBank ID: AF059183
• Promoter: T7
• Tags / Fusion Proteins:
  • GST (N terminal on backbone)
  • 6x His (N terminal on backbone)
  • S-tag (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: S-tag
• 3′ sequencing primer: T7 terminator

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Subcloned from a pET15b clone from O'Brien, J., Nguyen, H.B., and Mills, S.L. (2004). Cone photoreceptors in bass retina use two connexins to mediate electrical coupling. J Neurosci 24, 5632-5642.

Original cDNA clone is from O'Brien, J., Bruzzone, R., White, T.W., Al-Ubaidi, M.R., and Ripps, H. (1998). Cloning and expression of two related connexins from the perch retina define a distinct subgroup of the connexin family. J Neurosci 18, 7625-7637.