Oct4 단백질의 DNA - 결합 영역에 융합된 HIV - 1 integrase = HIV - 1 Integarse Fused to DNA - Binding Domain of Oct4 Protein

다국어 초록 (Multilingual Abstract)

Human immunodeficience virus type-1 (HIV-1) integrase (IN) mediates integration of viral DNA into the host cell genome which is an essential step during the life cycle of retroviruses. HIV-1 IN shows four enzymatic activities in vitro: non-specific DNA- binding, endonucleolytic (3'-processing), integration (strand transfer), and disintegration activities. Oct4 as a transcription factor recognizes and binds the specific sequences containing 5' -ATGCAAAT-3'. In order to study the specific binding and enzymatic activities of the integrase protein, we constructed the four different fusion proteins (OctIN1, OctIN2, INKOct1, INKOct1) using different combination of HIV-1 IN and Oct4 protein. The fusion proteins were overexpressed in Escherichia coli and purified by Ni-NTA and SP-sepharose columns. Analysis of the enzymatic activities showed that all the four fusion proteins has detectable endonucleolytic and integration activities, suggesting that the fusion proteins can be used to study derected integration and sequence specific DNA-binding of the integrase fusion protein.