profinity exacttm purification resin  (Bio-Rad)

Journal: Methods in Molecular Biology

Article Title: Heterologous Gene Expression in E.coli

doi: 10.1007/978-1-4939-6887-9

Figure Lengend Snippet: Fig. 1 Schematic representation of the Profinity eXact™ vectors. The Bio-Rad vectors pPAL7 and pPAL8 harbor only an N-terminal Profinity eXact™ tag (green) followed by a multiple cloning site (MCS, orange). The other vectors were constructed in-house and are based on the pET vector backbone (Novagen) to contain 6× His-tag (marked in red) with or without an additional expression enhancing tag (blue). The components of the expres- sion cassette are not drawn to scale. The MCS and the antibiotic resistance in the Profinity-pET based vectors differ from the ones in pPAL7 and pPAL8 (see Note 4)

Article Snippet: Mix the Profinity eXactTM affinity resin and transfer 2 mL of resin suspension (equivalent to 1 mL of settled beads) to the Bio-Rad Econo-Pac column.

Techniques: Cloning, Construct, Plasmid Preparation, Expressing

Journal: Methods in Molecular Biology

Article Title: Heterologous Gene Expression in E.coli

doi: 10.1007/978-1-4939-6887-9

Figure Lengend Snippet: Fig. 2 Comparison of protein expression using different solubility tags. The Water Soluble Chlorophyll Protein gene (WSCP, amino acids 12–190, NCBI XP_013613804.1) was cloned into each of the vectors listed in Fig. 1 (see Note 5). Expression of proteins was performed at 37 °C for 3 h. Cell pellets were pro- cessed in parallel, as described in the text. Analysis was performed using Bolt 4–12% Bis-Tris plus gel (Invitrogen). S- Soluble fraction, following cell lysis. E- Elution fraction following cleavage from the Profinity eXact™ tag. Arrow indi- cates position of the WSCP following cleavage. Asterisk indicates position of the full-length fusion protein in each of the soluble fraction

Article Snippet: Mix the Profinity eXactTM affinity resin and transfer 2 mL of resin suspension (equivalent to 1 mL of settled beads) to the Bio-Rad Econo-Pac column.

Techniques: Comparison, Expressing, Solubility, Clone Assay, Lysis