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.
pACYCDuet-1 is designed for the coexpression of two target genes. The vector encodes two multiple cloning sites (MCS) each of which is receded by a T7 promoter, lac
operator and ribosome binding site (rbs). The vector also carries the P15A replicon, lacI
gene and chloramphenicol resistance gene. This vector can be transformed into the same cell with pETDuet-1
, and pRSFDuet-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 ACYCDuetUP1 Primer
and DuetDOWN1 Primer
. Genes inserted into MCS-2 can be sequenced using the DuetUP2 Primer
and T7 Terminator Primer
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