A family of LIC vectors for high-throughput cloning and purification of proteins.
Fifteen related ligation-independent cloning vectors were constructed for high-throughput cloning and purification of proteins. The vectors encode a TEV protease site for removal of tags that facilitate pro tein purification (his-tag) or improve solubility (MBP, GST). Specialized vectors allow coexpression and copurification of interacting proteins, or in vivo removal of MBP by TVMV protease to improve screening and purification. All target genes and vectors are processed by the same protocols, which we describe here.
Eschenfeldt
William H
WH
Biosciences Division, Argonne National Laboratory, Lemont, IL, USA.
Lucy
Stols
S
Millard
Cynthia Sanville
CS
Joachimiak
Andrzej
A
Mark
I Donnelly
ID
D002477:Cells / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods; D010450:Endopeptidases / Q000235:genetics; D005822:Genetic Vectors; D016133:Polymerase Chain Reaction / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification*; D014170:Transformation, Genetic
D011994:Recombinant Proteins; D010450:Endopeptidases; C099488:TEV protease