pALEX, a dual-tag prokaryotic expression vector for the purification of full-length proteins.
pALEX, a prokaryotic expression vector, was constructed in which the multiple cloning site (MCS, polylinker) is flanked by sequences encoding glutathione S-transferase (GST) at the 5' end and a His6 residue tag at the 3' end. Open reading frames cloned into this vector can direct production of fusion proteins with GST at their N terminus and a His6 tag at their C terminus. This allows for the purification of full-size fusion proteins by a sequential two-step procedure on glutathione-agarose and Ni(2+)-agarose columns.
Panagiotidis
C A
CA
Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
Silverstein
S J
SJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics; D017874:Immediate-Early Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011387:Prokaryotic Cells; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification*; D044767:Ubiquitin-Protein Ligases
D017874:Immediate-Early Proteins; D011993:Recombinant Fusion Proteins; D044767:Ubiquitin-Protein Ligases; C051804:Vmw110 protein, Human herpesvirus 1; D005982:Glutathione Transferase