Coexpression of multiple target genes in E. coli
is advantageous for studying protein complexes. Coexpression often achieves optimal yield, solubility, and activity and may protect individual subunits from degradation. The Duet vectors are T7 promoter expression vectors, each designed to coexpress two target proteins in E. coli
. The Duet vectorscarry compatible replicons and antibiotic resistance markers and may be used together in appropriate host strains to coexpress up to eight proteins. Certain combinations of Duet vectors and pET or pETcoco vectors are also compatible for coexpression. The capability of Duet vectors to be cotransformed, propagated, and induced for robust target proteincoexpression makes them ideal for the analysis of protein complexes. The Duet vectors are designed with compatible replicons and drug resistance genes for effective propagation and maintenance of four plasmids in a single cell.
pETDuet-1 is designed for the coexpression of two target genes. The vector encodes two multiple cloning sites (MCS), each of which is preceded by a T7 promoter, lac
operator and ribosome binding site (rbs). The vector also carries the pBR322-derived ColE1 replicon, lacI
gene and ampicillin resistance gene. This vector can be transformed into the same cell with pACYCDuet-1
, and pRSFDuet-1
or pCOLADuet-1 in the appropriate host strain for the coexpression of up to eight target genes. Genes inserted into MCS-1 can be sequenced using the pETUpstream Primer
and DuetDOWN1 Primer
. Genes inserted into MCS-2 can be sequenced using the DuetUP2 Primer
and T7 Terminator Primer
. Commercial use of this Product requires a commercial license from EMD Millipore Corporation. Commercial use shall include but not be limited to (1) use of the Product or its components in manufacturing; (2) use of the Product or its components to provide a service, information, or data to others in exchange for consideration; (3) use of the Product or its components (or any derivatives thereof) for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the Product or its components, whether or not such Product or its components are resold for use in research. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of this Product.