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By Neamati N. (ed.)
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Extra resources for HIV-1 Integrase
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The sites of attack on the two target DNA strands are separated by five nucleotides in the case of HIV-1. (d) The product of DNA strand transfer is the integration intermediate in which the 30 ends of the viral DNA are joined to the 50 ends of the cleaved target DNA at the site of integration. The sites of joining on the two target DNA strands are separated by five nucleotides with a 50 protruding stagger in the case of HIV-1. The 50 ends of the viral DNA and the 30 ends of the target DNA are unjoined in the integration intermediate.
USA 1984, 81, 6461À6465. 34. ; Goff, S. P. Construction and analysis of deletion mutations in the pol gene of Moloney murine leukemia virus: A new viral function required for productive infection. Cell 1984, 37, 1043À1052. 35. Panganiban, A. ; Temin, H. M. The retrovirus pol gene encodes a product required for DNA integration: Identification of a retrovirus int locus. Proc. Natl. Acad. Sci. USA 1984, 81, 7885À7889. 36. Hippenmeyer, P. ; Grandgenett, D. P. Mutants of the Rous sarcoma virus reverse transcriptase gene are nondefective in early replication events.
Fractions were assayed for DNA binding by nitrocellulose filter binding using different radioactive-labeled DNAs as probes. The a and ab DNA polymerases and p12 (nucleocapsid, or NC) were identified along with an unknown DNA binding protein that eluted at a high salt concentration from the column. The elution profile of this unknown DNA binding protein was a lucky break for us. This protein turned out to be p32, and its purification using phosphocellulose chromatography was employed in the laboratory for many years.