While multi-protein expression remains a formidable task and generating sufficiently soluble protein complexes at times requires some tweaking, such challenges now become easier with MultiColi™ a versatile and convenient Acceptor-Donor vector system for manual or automated recombineering of genes destined for expression in E. coli. The MultiColi™ technology is the first fully automatable system for protein complex production in E.coli. Originally developed as part of EMBL’s ACEMBL pipeline, which is a centerpiece of the European Commission Framework Programme 7 project ComplexINC, MultiColi™ sets new standards for multi-protein expression in this versatile laboratory host organism.
MultiColi™ transfer vectors are fully synthetic, comprising only functional DNA, and as a result are small and easy to handle.
How MultiColi™ Works:
- Left) Design and clone your gene(s) of interest (GOIs) into MultiColi™ acceptor and donor vectors
- Center) Match and mix your GOIs and then (re)combine them into one construct via Cre-Lox recombination, and select your construct using unique combination of antibiotic markers
- Right) Transform E. coli cells with composite GOI vector. Expression follows via induction.
Resulting multiprotein expression product can be used for biochemical analysis, structural biology, drug discovery etc.
References and Downloads:
Click here to download Bieniossek et al 2009. Nature Methods.
Click here to download Nie et al 2009. Nature Protocols.
Click here to download MultiColi™ User Manual v5.1
Click here for DNA vector sequences
Click here to download Cre-ACEMBLER User Manual 3.0
Click here to download Cre-ACEMBLER Short Description
Click here to download Cre-ACEMBLER for Windows
Click here to download Cre-ACEMBLER for Linux