Carteau S, Mouscadet J F, Goulaouic H, Subra F, Auclair C
Laboratoire de Pharmacologie Moléculaire, CNRS URA 147, INSERM U 140, Institut Gustave Roussy, Villejuif, France.
Arch Biochem Biophys. 1993 Feb 1;300(2):756-60. doi: 10.1006/abbi.1993.1105.
An obligatory step of retroviral growth is the integration of a DNA copy of the viral RNA into the genomic DNA of the host. Recombinant human immunodeficiency virus type I (HIV-1) integrase (IN) expressed in Escherichia coli efficiently catalyzes the overall in vitro integration reaction, namely, the processing of the long terminal repeat (LTR) ends and the strand transfer reaction. Using the 3' end of synthetic oligonucleotides which match the termini of HIV-1 LTRs as substrate and supercoiled pSP65 DNA as the target, we describe an assay that is suitable for the enzymatic analysis of the integration and for testing candidate inhibitors of HIV IN protein.
逆转录病毒生长的一个必要步骤是将病毒RNA的DNA拷贝整合到宿主的基因组DNA中。在大肠杆菌中表达的重组人免疫缺陷病毒I型（HIV-1）整合酶（IN）能有效地催化整个体外整合反应，即长末端重复序列（LTR）末端的加工和链转移反应。我们以与HIV-1 LTR末端匹配的合成寡核苷酸的3'末端为底物，以超螺旋pSP65 DNA为靶标，描述了一种适用于整合酶促分析和测试HIV IN蛋白候选抑制剂的检测方法。
