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首页> 外文期刊>Molecular and Cellular Biology >Proviral sequences that restrict retroviral expression in mouse embryonal carcinoma cells.
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Proviral sequences that restrict retroviral expression in mouse embryonal carcinoma cells.

机译：限制逆转录病毒在小鼠胚胎癌细胞中表达的原病毒序列。

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摘要

Embryonal carcinoma (EC) cells are nonpermissive for retrovirus replication. Restriction of retroviral expression in EC cells was studied by using DNA transfection techniques. To investigate the activity of the Moloney murine leukemia virus (M-MuLV)enhancer and promoter sequences, the M-MuLV long terminal repeat and the defined long terminal repeat deletions were linked to neo structural gene sequences that encode resistance to the neomycin analog G418. Transient expression data and drug resistance frequencies support the findings that the M-MuLV enhancer is not active in EC cells but that promoter sequences are functional. In addition, a proviral DNA fragment that encodes the leader RNA sequence of a M-MuLV recombinant retrovirus was found to restrict expression specifically in EC cells. Deletion analysis of the leader fragment localized the inhibitory sequences to a region that spans the M-MuLV tRNA primer binding site. It is not known whether restriction occurs at a transcriptional or posttranscriptional level, but steady-state RNA levels in transient expression assays were significantly reduced.
机译：胚胎癌细胞（EC）不允许逆转录病毒复制。通过使用DNA转染技术研究了EC细胞中逆转录病毒表达的限制。为了研究莫洛尼氏鼠白血病病毒（M-MuLV）增强子和启动子序列的活性，将M-MuLV长末端重复序列和定义的长末端重复序列缺失与编码对新霉素类似物G418的抗性的新结构基因序列相关联。瞬时表达数据和耐药频率支持以下发现：M-MuLV增强子在EC细胞中不活跃，但启动子序列具有功能。另外，发现编码M-MuLV重组逆转录病毒的前导RNA序列的前病毒DNA片段特异性地限制了在EC细胞中的表达。前导片段的缺失分析将抑制序列定位在跨越M-MuLV tRNA引物结合位点的区域。尚不清楚限制是发生在转录水平还是转录后水平，但是瞬时表达测定法中的稳态RNA水平显着降低。

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