首页> 美国卫生研究院文献>Journal of Virology >Regulated expression of the feline panleukopenia virus P38 promoter on extrachromosomal FPV/EBV chimeric plasmids.
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Regulated expression of the feline panleukopenia virus P38 promoter on extrachromosomal FPV/EBV chimeric plasmids.

机译:猫泛白细胞减少症病毒P38启动子在染色体外FPV / EBV嵌合质粒上的表达调控。

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摘要

Feline panleukopenia virus/Epstein-Barr virus (FPV/EBV) chimeric expression plasmids were constructed to study regulation of the structural protein gene of the parvovirus, FPV, in a homologous cell culture system. Detection and quantitation of activity from the native FPV promoter, P38, was facilitated by fusing the Escherichia coli lacZ gene with the FPV structural protein gene. Feline cell lines which stably maintained these plasmids extrachromosomally were established. Constitutive beta-galactosidase activity was low but increased up to 40-fold after infection with FPV. Expression of beta-galactosidase was only detected when the FPV/lacZ gene was oriented in the same transcriptional direction as the Epstein-Barr virus gene coding for EBNA-1. When a small open reading frame upstream of the FPV/lacZ initiation codon was deleted, beta-galactosidase expression increased another 4.7- to 26-fold. These changes in beta-galactosidase activity indicate that expression of the FPV structural protein gene is regulated both transcriptionally and posttranscriptionally.
机译:构建猫泛白细胞减少病毒/爱泼斯坦-巴尔病毒(FPV / EBV)嵌合表达质粒,以研究在同源细胞培养系统中细小病毒FPV的结构蛋白基因的调控。通过将大肠杆菌lacZ基因与FPV结构蛋白基因融合,可以方便地检测和定量来自天然FPV启动子P38的活性。建立了稳定地在染色体外维持这些质粒的猫细胞系。 FPV感染后,本构性β-半乳糖苷酶活性较低,但最高可增加40倍。仅当FPV / lacZ基因的方向与编码EBNA-1的爱泼斯坦-巴尔病毒基因的转录方向相同时,才能检测到β-半乳糖苷酶的表达。当FPV / lacZ起始密码子上游的一个小的开放阅读框被删除时,β-半乳糖苷酶的表达又增加了4.7到26倍。 β-半乳糖苷酶活性的这些变化表明FPV结构蛋白基因的表达在转录和转录后均受到调节。

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