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Comparative studies of recombinant reverse transcriptases from HIV-2 and SIV.




 

Int Conf AIDS. 1992 Jul 19-24;8(3):38 (abstract no. PuA 6166). Unique

OBJECTIVE: Production of HIV-2 and SIV RTs for comparative biochemical and structural analysis. METHODS: The coding sequences for HIV-2 and SIV RTs were obtained by PCR from genomic clones pROD35#2 and SIVmac32H(pj5) respectively. The 3' PCR primer incorporated sequences to mutate the C-termini to GLU-GLU-PHE. This sequence is recognised by a Mab (YL1/2) which was used in immobilised form to purify these RTs expressed in E.coli. Purified RTs were characterised both kinetically as well as by reaction with antibodies (YL1/2 and an antipeptide antibody to an SIV C-terminal peptide). RESULTS AND CONCLUSIONS: Both HIV-2 and SIV RTs were purified in a single step using the antibody column, and were mainly observed as 68K polypeptides with small amounts of 55K. A C-terminal fragment of apparent Mr 40K was seen for HIV-2 but not SIV RT. The reaction of the 55K form with the SIV peptide antibody indicated the cleavage site for the protease in both HIV-2 and SIV RTs could be further towards the C-terminus than in HIV-1 RT.

Cloning, Molecular Comparative Study HIV-2/*ENZYMOLOGY/GENETICS Molecular Weight Polymerase Chain Reaction Recombinant Proteins/ISOLATION & PURIF/METABOLISM RNA-Directed DNA Polymerase/GENETICS/ISOLATION & PURIF/ *METABOLISM SIV/*ENZYMOLOGY/GENETICS ABSTRACT



 




Information in this article was accurate in December 30, 1992. The state of the art may have changed since the publication date. This material is designed to support, not replace, the relationship that exists between you and your doctor. Always discuss treatment options with a doctor who specializes in treating HIV.