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

Abstract:

: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 enzyme. The p15 RNaseH domain has been purified using this method and shown to have low enzyme activity compared to the parental p66 subunit.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Stammers DK,Tisdale M,Court S,Parmar V,Bradley C,Ross CK

doi

10.1016/0014-5793(91)80613-8

subject

Has Abstract

pub_date

1991-06-03 00:00:00

pages

298-302

issue

2

eissn

0014-5793

issn

1873-3468

pii

0014-5793(91)80613-8

journal_volume

283

pub_type

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