An efficient expression, purification and immunodetection system for recombinant gene products

Biotechniques. 1994 Jul;17(1):94, 96, 98-9.


We describe a modification of the mammalian expression vector pRc/CMV, which drives expression of inserted genes from either the human cytomegalovirus (CMV) immediate-early promoter or the bacteriophage T7 RNA polymerase promoter. The modification is designed to allow expression, simple purification and specific immunodetection of recombinant fusion proteins. The modified plasmid, termed pTag/CMV-neo, encodes a Kozak consensus ribosome-binding site (RBS) and a 30-amino acid fusion tag peptide. This peptide consists of a metal ion-binding site, (His)6, for single-step affinity purification using Ni(2+)-chelating resin and a multi-purpose HIV-1-derived peptide (p18HIV). This viral epitope can be used to identify, detect and characterize target fusion proteins in conjunction with a specific monoclonal antibody H902 that does not display cross-reactivity with cellular proteins. The H902 production hybridoma cell line is reagent #521 from the NIH AIDS Research and Reference Program.

Publication types

  • Research Support, U.S. Gov't, P.H.S.
  • Technical Report

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cytomegalovirus / genetics
  • DNA-Directed RNA Polymerases / genetics
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
  • Molecular Sequence Data
  • Plasmids
  • Promoter Regions, Genetic
  • Proto-Oncogene Proteins / biosynthesis
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / isolation & purification
  • Viral Proteins


  • Proto-Oncogene Proteins
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases