Coulombe J, Avis Y, Gray D A
Department of Biochemistry, University of Ottawa, Ontario, Canada.
J Virol. 1996 Oct;70(10):6810-5. doi: 10.1128/JVI.70.10.6810-6815.1996.
A promoter-trap retrovirus has been constructed in which a promoterless polyomavirus middle T antigen gene was inserted in the U3 region of the long terminal repeat of a replication-competent Moloney murine leukemia virus. The resulting virus, designated PyT, was used to infect mouse mammary glands in situ. As expected, mammary tumors appeared in some infected animals. These tumors were found to contain PyT proviruses of the predicted structure. From one such tumor, the PyT provirus and surrounding sequences from the integration site were cloned. The provirus was found to have integrated adjacent to the promoter of a novel mouse gene (TRAP1) that was expressed at low levels in various mouse tissues. These data show that the PyT retrovirus provides a sensitive means of detecting active promoters in vivo.
构建了一种启动子捕获逆转录病毒，其中无启动子的多瘤病毒中间T抗原基因插入到具有复制能力的莫洛尼鼠白血病病毒长末端重复序列的U3区域。所得病毒命名为PyT，用于原位感染小鼠乳腺。如预期的那样，一些受感染动物出现了乳腺肿瘤。这些肿瘤被发现含有预测结构的PyT前病毒。从一个这样的肿瘤中，克隆了PyT前病毒及其整合位点周围的序列。发现前病毒整合在一个新的小鼠基因（TRAP1）启动子附近，该基因在各种小鼠组织中低水平表达。这些数据表明，PyT逆转录病毒提供了一种在体内检测活性启动子的灵敏方法。
