pEAT8-137 M1V
(Plasmid #173807)

• Purpose: Expression and purification of mature (no signal sequence) human alpha1-antitrypsin wildtype variant M1V (UniProt P01009) from E. coli BL21(DE3) cells
• Depositing Lab: Anne Gershenson
• Publication: Krishnan et al Methods Mol Biol. 2017;1639:195-209. doi: 10.1007/978-1-4939-7163-3_19.

Backbone

• Vector backbone: pET-8C
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: E coli BL21(DE3) can be used for expression. Note that the temperature for protein induction for soluble expression is 30 C (see PubMed 28752459).
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: SERPiNA1
• Alt name: alpha1-antitrypsin M1V
• Species: H. sapiens (human)
• Mutation: E1M Otherwise this plasmid encodes the most common wildtype form of human alpha1-antitrypsin called M1V. The sequence is that of the mature protien (no signal seqquence) in UniProt entry P01009
• Entrez Gene: SERPINA1 (a.k.a. A1A, A1AT, AAT, PI, PI1, PRO2275, alpha1AT, nNIF)
• Promoter: T7

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pubmed 7696306 (original cloning)

Terms and Licenses

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

Depositor Comments

For details on mutating an internal initiation site in the SERPIINA1 gene (changing the pEAT8 plasmid to pEAT8-137) see: Laska, M. E. (2001) The effect of dissolved oxygen on recombinant protein degradation in Escherichia coli, Thesis, Massachusetts Institute of Technology, Cambridge, MA