Automated unrestricted multigene recombineering for multiprotein complex production.
Structural and functional studies of many multiprotein complexes depend on recombinant-protein overexpression. Rapid revision of expression experiments and diversification of the complexes are often crucial for success of these projects; therefore, automation is increasingly indispensable. We introduce Acembl, a versatile and automatable system for protein-complex expression in Escherichia coli that uses recombineering to facilitate multigene assembly and diversification. We demonstrated protein-complex expression using Acembl, including production of the complete prokaryotic holotranslocon.
European Molecular Biology Laboratory, Grenoble Outstation, Grenoble, France.
Christoph
C
Bieniossek
Yan
Y
Nie
Daniel
D
Frey
Natacha
N
Olieric
Christiane
C
Schaffitzel
Ian
I
Collinson
Christophe
C
Romier
Philipp
P
Berger
Timothy J
TJ
Richmond
Michel O
MO
Steinmetz
Imre
I
Berger
D004926:Escherichia coli / Q000502:physiology*; D005810:Multigene Family / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis*
D011994:Recombinant Proteins