PEG-Actinin-M-sstFRET
(Plasmid #61100)

• Purpose: Expression of α-actinin with internal sstFRET inserted to measure cytoskeletal stress in vivo
• Depositing Lab: Fred Sachs
• Depositing Organization: University at Buffalo (SUNY Buffalo)
  Located in United States of America
• Publication: Meng et al J Cell Sci. 2011 Jan 15;124(Pt 2):261-9. doi: 10.1242/jcs.071928. Epub 2010 Dec 20.
• Citations: 1 Article

Backbone

• Vector backbone: pEYFP–C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• Total vector size (bp): 8432
• Modifications to backbone: To create chimeric gene constructs of α-actinin, we subcloned the actinin gene into pEYFP–C1 vector, where the YFP gene was removed beforehand. The restriction enzyme sites introduced into PCR products for subcloning were NheI-Actinin-KpnI.
• Vector type: Mammalian Expression
• 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: Unknown

Gene/Insert

• Gene/Insert name: α-actinin-sstFRET
• Mutation: sstFRET inserted at position 300
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: CMV Forward
• 3′ sequencing primer: SV40 poly A Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pEYFP–C1 Venus and pECFP–C1 Cerulean plasmids were generous gifts from David W. Piston. sstFRET were inserted into actinin at position 300, which was located in the linker domain between first and second spectrin repeat domains by restriction sites AgeI and NotI. The restriction enzyme sites were introduced into the host proteins using the site-directed mutagenesis kit from Stratagene (La Jolla, CA) with the host protein amino acid unchanged. An 8-histidine tag followed by TAA stop codon was inserted in front of the NotI site to make sure that the tag was located in the C-terminal and well-exposed to solution.

Terms and Licenses

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

DNA (Catalog # 61100-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 61100-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.