Institute for Cancer Research, Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111, USA.
Methods. 2009 Apr;47(4):243-8. doi: 10.1016/j.ymeth.2008.10.024. Epub 2008 Nov 14.
Oligonucleotide assays have been invaluable for elucidation of the molecular mechanisms of retroviral integrases. A suite of rapid and sensitive fluorescence assays to measure the DNA binding, processing, and joining activities of integrase (IN) is described here. The assays are especially useful for characterizing the major activities of the enzyme, and for handling large numbers of samples efficiently. They can greatly facilitate further biochemical and structural analyses for HIV-1 and other IN proteins. The assays can also be adapted for moderate-high throughput testing of various inhibitory compounds.
寡核苷酸检测在阐明逆转录病毒整合酶的分子机制方面具有不可估量的价值。本文描述了一套快速灵敏的荧光检测法,用于测量整合酶(IN)的 DNA 结合、加工和连接活性。这些检测法特别适用于描述酶的主要活性,以及高效处理大量样本。它们可以极大地促进对 HIV-1 和其他 IN 蛋白的进一步生化和结构分析。这些检测法也可以适应于各种抑制化合物的中高通量测试。