Vmax™ Express is a novel, fast-growing bacterial strain designed and optimized for high-level recombinant protein expression. This rationally engineered, next-generation prokaryotic protein expression system can serve as a replacement for slow growing E. coli systems that are prone to low yields and the expression of proteins as insoluble inclusion bodies. Vmax™ cells are derived from the marine microorganism, Vibrio natriegens. This gram-negative, non-pathogenic bacterium has a doubling time twice that of E. coli and produces 3-4 times the biomass. Coupled with a robust transcription and translation systems to support this rapid growth rate, Vmax™ Express generates greater amounts of biomass and more recombinant protein per liter faster.
With a protein expression workflow similar to E. coli, Vmax™ Express is compatible with plasmids and antibiotics commonly used with bacterial expression systems such as E. coli BL21(DE3). The Vmax™ Express workflow, fast growth, and ability to express a wide range of proteins make it an ideal host for routine protein expression or for proteins that do not express well in E. coli. Additionally, Vmax™ Express can help overcome common recombinant protein expression challenges such as low yields, or the expression of insoluble, or inactive proteins. Built with a tightly controlled, IPTG-inducible T7 promoter system, Vmax™ Express cells can be cultured using routine growth medium, as well as commercial auto-induction media, or our Vmax™ Enriched Growth Media (recommended for rapid growth and greatest accumulation of biomass).
In contrast to E. coli, Vmax™ Express cells grow rapidly at both 30°C and 37°C and tolerate the induction of protein expression over a wide range of starting ODs and IPTG concentrations. To maximize the amount of soluble protein produced In addition, Vmax™ Express cells can be incubated for up to 24 hours post induction. This flexible, high-yield protein expression workflow and rapid doubling time allows you to go from transformed cells or a glycerol stock to a large-scale culture ready for analysis or protein purification in as few as three standard workdays compared to four days with E. coli.
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|Protein Expression Strain Performance Attribute||E. Coli Strains||Vmax™ Express|
|Compatible with standard plasmid origins of replication (e.g. pMB1, ColE1, pUC, p15A)||✔||✔|
|Able to use common inexpensive growth media||✔||✔|
|Plasmid selection uses common antibiotic resistance markers (Amp, Tet, Kan, Cm)||✔||✔|
|Tightly controlled IPTG Inducible T7 transcription system||✔||✔|
|Doubling time of ~14 minutes||✖||✔|
|Biomass >14 OD after 24 hours growth||✖||✔|
|Flexibility to induce expression at range of OD600||✖||✔|
|Rapid growth at both 30° and 37°C*||✖||✔|
|Culture ready for protein purification and analysis in 3 days||✖||✔|
|Vmax™ Enriched Growth Media||Liquid Culture||Growth||Growth||Growth||++++|
|Enhanced 2xYT Media||Liquid Culture||Growth||Growth||Growth||+++ Recommended|
|Brain Heart Infusion Broth + v2 salts‡||Liquid Culture||Growth||Growth||Growth||++|
|LB-Miller†||Liquid Culture||Growth||Growth||Supplement with v2 salt‡||++|
|Terrific broth||Liquid Culture||Not tested||Growth||Growth||+|
|MagicMedia™||Liquid Culture||Not tested||Growth||Growth||+|
|LB-Miller†||Agar Plate||Slow Growth||Growth||Growth||+++ Recommended|
|Brain Heart Infusion agar + v2 salts‡||Agar Plate||Growth||Growth||Growth||++|
A variety of commonly used plasmid origins of replication and antibiotic selection markers are compatible with Vmax™ Express. This allows the use of existing expression constructs for recombinant protein expression. After transformation or recovery from a glycerol stock, Vmax™ Express cells can be plated on nutrient agar with appropriate antibiotics. The tables below provide the growth conditions and suggested antibiotic concentrations for both solid and liquid media.
|Plasmid Origin of Replication||Plasmid Backbone||Growth Plate Recommendation|
|pMB1||pBR322 pET vectors||Recover at 30 or 37°C
|Recover at 30 or 37°C
|pUC||pUC19||If using Kan selection, recover at 30°C only.|
|p15A||pACYC184||Recover at 30 or 37°C
|Antibiotic Marker||Concentration (µg/mL) for Solid Media||Concentration (µg/mL) for Liquid Culture|
|Ampicillin||10 - 50 μg/mL||50 - 25 μg/mL|
|Carbenicillin||2 - 25 μg/mL*||5 - 100 μg/mL|
|Kanamycin||100 μg/mL||200 μg/mL|
|Chloramphenicol||5 - 12.5 μg/mL||12.5 - 25 μg/mL|
*Vmax™ cells are more sensitive to carbenicillin than E. coli, whereas ampicillin sensitivity is similar to E. coli. When using solid media, concentrations above 12.5 μg/mL typically do not decrease transformation efficiency. However, higher antibiotic concentrations tend to result in colony size heterogeneity.
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