A high-throughput, non-radioactive microtiter plate assay for activity of the human immunodeficiency virus integrase protein
Nucleic Acids ResearchPublished 1 January 1994
Cornelis Vink, Martin S. Banks, Richard C. Bethell, Ronald H.A. PIasterk
Citations35
SJR quartileQ1
SJR score7.78
SNIP4.71
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TL;DR
The development of an efficient non-radioactive assay for IN activity which is carried out in a single well of a microtiter plate is described.
Abstract
Cornelis Vink, Martin Banks, Richard Bethell, Ronald H.A. PIasterk; A high-throughput, non-radioactive microtiter plate assay for activity of the human imm
Keywords
Immunology and MicrobiologyMedicine
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It is found that the DNA-binding region resides between amino acids 200 and 270 of the 288-residues HIV-1 IN protein, and the catalytic domain of the protein was mapped between amino acid 50 and 194.
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261 Citations1992Robert L. LaFemina, Christine L. Schneider +6 more
A double substitution of aspartic acid and glutamine for valine and glutamic acid at positions 151 and 152 within the D,D(35)E motif region rendered the integrase protein inactive for all of its functions, yielding a mutant virus that was incapable of productively infecting human T-lymphoid cells in culture.
Journal of VirologyIntegration is not necessary for expression of human immunodeficiency virus type 1 protein products
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HIV-1 mutants which were deleted in the integrase/endonuclease gene and which were unable to establish an integrated form of the virus were constructed and were fully competent templates for HIV-1 core and envelope antigen production.
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The microtiter plate assay is designed with the specific aim of developing a high volume, non-radioisotopic assay for HTV-1 integrase which uses unique oligonucleotides to represent the LTR donor and target DNA substrates.
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111 Citations1991Robert Craigie, Kiyoshi Mizuuchi +2 more
This work presents a simple assay system that is suitable for screening potential inhibitors of HIV DNA integration, and since each step of the assay can be carried out in the wells of microtiter plates, large numbers of reactions can be processed simultaneously.
