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首页> 外文期刊>Journal of Virology >High-frequency deletion in recovered retrovirus vectors containing exogenous DNA with promoters.
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High-frequency deletion in recovered retrovirus vectors containing exogenous DNA with promoters.

机译：恢复逆转录病毒载体的高频缺失含有与促进剂的外源性DNA。

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We previously described infectious retrovirus vectors constructed from spleen necrosis virus which contain the herpes simplex virus thymidine kinase gene and the mouse alpha-globin gene (K. Shimotohno and H. M. Temin, Nature [London] 299:255-268, 1982). In the present study we report that when TK- chicken cells infected with a virus containing the mouse alpha-globin promoter and other 5' noncoding sequences in addition to the alpha-globin coding sequences were selected for thymidine kinase (TK) activity, all virus-producing TK+ cell clones shed virus with a deletion. These deletions were of different sizes and included the mouse alpha-globin coding sequences and the mouse alpha-globin transcriptional promoter. One of the deleted viruses was molecularly cloned. DNA sequencing showed that the deleted sequences are flanked by a short direct repeat. This deleted virus was also shown to have an advantage over the nondeleted parent both in multiplication and in its specific TK-transforming unit titer. In contrast to the results described above, TK+ cell clones established with viruses that contained only the coding sequences from the mouse alpha-globin gene did not delete and were stable over many cell passages. The implications of the high-frequency deletion of the viruses with internal promoters are discussed in terms of the evolution of retroviruses and the construction of retrovirus vectors.
机译：我们以前描述了由脾脏坏死病毒构成的传染病病毒载体，该病毒含有疱疹病毒胸苷激酶基因和小鼠α-珠蛋白基因（K.Shimotohno和H. Temin，Nature [伦敦] 299：255-268,1982）。在本研究中，我们认为，除了含有小鼠α-珠蛋白启动子和其他5'非编码序列的病毒的TK-鸡细胞选择除了α-珠蛋白编码序列外，是否选择胸苷激酶（TK）活性，所有病毒 - 用缺失发挥Tk +细胞克隆病毒。这些缺失具有不同的尺寸并包括小鼠α-珠蛋白编码序列和小鼠α-珠蛋白转录启动子。其中一种缺失的病毒被分子克隆。 DNA测序表明缺失的序列侧重于短直接重复。该缺失的病毒也显示在乘法和其特定TK变换单元滴度上的非勃起的父级上具有优于非塞子父母。与上述结果相反，与仅包含来自小鼠α-珠蛋白基因的编码序列的病毒建立的TK +细胞克隆并未删除并且在许多细胞通道上稳定。在逆转录病毒的演变和逆转录病毒载体的构建方面，讨论了高频缺失病毒与内部启动子的影响。

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