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首页> 外文期刊>Molecular and Cellular Biology >Replication-defective chimeric helper proviruses and factors affecting generation of competent virus: expression of Moloney murine leukemia virus structural genes via the metallothionein promoter.
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Replication-defective chimeric helper proviruses and factors affecting generation of competent virus: expression of Moloney murine leukemia virus structural genes via the metallothionein promoter.

机译：复制缺陷的嵌合辅助促进助理潜水和影响竞争性病毒产生的因素：Moloney鼠白血病病毒结构基因通过金属硫蛋白促进剂的表达。

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Two chimeric helper proviruses were derived from the provirus of the ecotropic Moloney murine leukemia virus by replacing the 5'long terminal repeat and adjacent proviral sequences with the mouse metallothionein I promoter. One of these chimeric proviruses was designed to express the gag-pol genes of the virus, whereas the other was designed to express only the env gene. When transfected into NIH 3T3 cells, these helper proviruses failed to generate competent virus but did express Zn2+-inducible trans-acting viral functions needed to assemble infectious vectors. One helper cell line (clone 32) supported vector assembly at levels comparable to those supported by the Psi-2 and PA317 cell lines transfected with the same vector. Defective proviruses which carry the neomycin phosphotransferase gene and which lack overlapping sequence homology with the 5' end of the chimeric helper proviruses could be transfected into the helper cell line without generation of replication-competent virus. Mass cultures of transfected helper cells produced titers of about 10(4) G418r CFU/ml, whereas individual clones produced titers between 0 and 2.6 X 10(4) CFU/ml. In contrast, defective proviruses which share homologous overlapping viral sequences with the 5' end of the chimeric helper proviruses readily generated infectious virus when transfected into the helper cell line. The deletion of multiple cis-acting functions from the helper provirus and elimination of sequence homology overlapping at the 5' ends of helper and vector proviruses both contribute to the increased genetic stability of this system.
机译：通过用小鼠金属硫蛋白I启动子替换5'Long末端重复和相邻的荧光序列来源于生态摩洛狼白血病病毒的两种嵌合辅助潜水术。这些嵌合潜品之一被设计成表达病毒的GAG-POL基因，而另一个被设计用于仅表达ENV基因。当转染到NIH 3T3细胞中时，这些辅助潜水术未能产生态度病毒，但表达了组装传染性载体所需的表达Zn2 +--uduciby转发病毒功能。一个辅助细胞系（克隆32）支撑载体组件以与用相同载体转染的PSI-2和PA317细胞系支持的水平相当的水平。携带新霉素磷酸转移酶基因的缺陷潜水术并缺乏与嵌合辅助潜水潜艇的5'末端的重叠序列同源物体可以转染到辅助细胞系中而不产生复制态病毒。转染的辅助细胞的质量培养物产生约10（4）G418R CFU / mL的滴度，而单个克隆产生0至2.6×10（4）CFU / mL的滴度。相反，缺陷的潜水术与嵌合辅助助手的5'末端分享同源的重叠病毒序列，当转染到辅助细胞系中时容易产生传染性病毒。从辅助概况和消除助手和载体潜水术的5'末端消除枢孔概况和消除序列同源性的多个顺式同源功能均有助于该系统的遗传稳定性增加。

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