With the pEU Vector Set, we are providing dedicated expression vectors for use in the wheat germ cellfree protein expression system. This includes vectors for working with the GST-, His, or DYKDDDDK-affinity tag as well as the positive control vector pEU-E01-DHFR for testing your reaction conditions. Each vector has an SP6 RNA polymerase promoter to drive RNA expression followed by the E01 enhancer. The E01 enhancer is required to initiate the translation reaction. The coding region for the protein of interest can be cloned into any of the restriction sites within the multi cloning site of the vectors. However, we advise to use the closest possible restriction site near the E01 enhancer to assure effective initiation of translation. The SP6 RNA polymerase does not require any terminator sequence to stop transcription.
Inserts can be cloned into the vectors using standard methods for DNA recombination. Please assure during vector design that your insert encodes for a proper starting methionine and has an in frame stop codon at the end; pEU vectors do not provide stop codons after the multi cloning site. Assure further that inserts are in frame when using a vector encoding for an affinity tag at the N terminus. A starting methionine may not be required when cloning in frame with an affinity tag at the N terminus. Similarly, do not include any stop codon at the end of your insert when cloning in frame with an affinity tag at the C terminus.
All our vectors have an ampicillin resistance marker. After transformation bacteria can be grown on Lysogeny Broth (LB) medium with added ampicillin at a concentration of 100 μg/mL. The vectors are highcopy vectors and should commonly give good DNA yields using regular plasmid DNA purification kits. Confirm DNA purity before use in a protein expression experiment. It may be necessary to further purify the plasmid DNA by phenol/chloroform extraction if no protein expression is observed.
pEU vector is an expression vector that has been highly optimized to maximize synthetic efficiency through a wheat germ cell-free protein expression system. By introducing bacteriophage SP6 into the transcriptional promoter and a completely artificial E01 sequence selected from a randomized nucleic acid sequence pool into the translational enhancer, an mRNA with high translational activity can be produced, and protein synthesis efficiency can be maximized. In addition to the vector outlined above, our company also offers vectors with added His-tag, GST-tag and DYKDDDDK-tag that are suited to affinity purification. You can choose which one you need depending on the situation.
Exclusively for your first-time purchase of kits/reagents, this set will be included free of charge.
*Please note: For your second and subsequent orders, purchase will be required.
Please contact us at for specific academic pricing.