Martín F, Jean-Mairet Y, Talavera A
Centro de Biología Molecular Severo Ochoa (CSIC-UAM), Cantoblanco, España.
Microbiologia. 1996 Mar;12(1):43-50.
We have studied the effect that exerts on the Moloney murine leukemia virus (Mo-MLV) viral cycle the deletion of the 13 bases that constitute the inverted repeat (IR) present at the external U3 region of the pre-proviral DNA. Whereas supernatants of wild type- and modified virus-producing cells contained similar amounts of viral particles, the deleted viruses showed a 100 to 1000-fold decreased infectivity. To determine how the deletion interfered with the infective capacity of the virus, different steps of the viral cycle were studied using deleted viruses. The deletion affected neither reverse transcription nor the entry of the DNA into the cell nucleus; however, the integration of the pre-proviral DNA into the host genome was reduced to undetectable levels. These results open a pathway to the construction of Mo-MLV-derived retroviral vectors for gene targeting purposes.
我们研究了前病毒DNA外部U3区域存在的构成反向重复序列（IR）的13个碱基缺失对莫洛尼鼠白血病病毒（Mo-MLV）病毒周期的影响。野生型和修饰病毒产生细胞的上清液中含有相似数量的病毒颗粒，但缺失病毒的感染性降低了100至1000倍。为了确定缺失如何干扰病毒的感染能力，使用缺失病毒研究了病毒周期的不同步骤。该缺失既不影响逆转录，也不影响DNA进入细胞核；然而，前病毒DNA整合到宿主基因组中的水平降低到无法检测的程度。这些结果为构建用于基因靶向目的的Mo-MLV衍生逆转录病毒载体开辟了一条途径。
