Rapid purification and characterisation of HIV-1 reverse transcriptase and RNaseH engineered to incorporate a C-terminal tripeptide alpha-tubulin epitope.

Journal article

The C-termini of p66 and p51 forms of HIV-1 reverse transcriptase have been engineered to contain a Glu-Glu-Phe sequence recognized by a monoclonal antibody to alpha-tubulin, YL1/2. Mutated RTs were purified in a single step using peptide elution from columns of immobilized YL1/2. The known sequence requirements of the YL1/2 epitope are consistent with protein eluting from the column with an intact C-terminus. Kinetic parameters of these mutated RTs are essentially unchanged from wild-type en...

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Authors: Stammers, D; Tisdale, M; Court, S; Parmar, V; Bradley, C

• Publication status: Published
• Journal: FEBS letters
• Volume: 283
• Issue: 2
• Pages: 298-302
• Publication date: 1991-06-01
• DOI: 10.1016/0014-5793(91)80613-8
• EISSN: 1873-3468
• ISSN: 0014-5793
• Language: English
• Keywords: Molecular Weight; Amino Acid Sequence; Viral Structural Proteins; HIV Protease; Epitopes; Electrophoresis, Polyacrylamide Gel; Endoribonucleases; Recombinant Proteins; Genes, Viral; Mutagenesis, Site-Directed; Ribonuclease H; Restriction Mapping; RNA-Directed DNA Polymerase; Tubulin; Genetic Engineering; Molecular Sequence Data; Polymerase Chain Reaction; Chromatography, Affinity; Blotting, Western; HIV-1