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十六烷基三乙基溴化铵对家蚕核型多角体病毒复制的影响。

Effects of cetyltriethylammonium bromide on the replication of Bombyx mori nucleopolyhedrovirus.

机构信息

Key Laboratory of Silkworm Biotechnology, Ministry of Agriculture, Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu, China.

出版信息

Cytotechnology. 2002 May;39(1):27-35. doi: 10.1023/A:1022496703875.

Abstract

An experimental study was undertaken to quantify the effects of cetyltriethylammonium bromide (CTAB) on the replication of Bombyx mori nucleopolyhedrovirus (BmNPV) and the transcriptionalactivity of BmNPV ie-1 promoter. The results demonstrated that the budded virus (BV) titer rose about 3.7-fold by adding CTAB to the culture media up to 0.1 mu g ml(-1) in infected Bm-N cells with a wild-type BmNPV. The transient expression level of luciferase driven by BmNPV ie-1 promoter was enhanced by more than 3-fold in the presence of 0.1 mu g ml(-1) of CTAB in uninfected insect cells via a transient expression system. Contrary to the rise in BV titer, the polyhedra inside the nucleus of infected cells dropped linearly from 4.0 x 10(6) ml(-1) down to 2.1 x 10(6) ml(-1) with in a range of CTAB concentrations from 0 to 0.25 mu g ml(-1). The same trend in expression level of beta -galactosidase or phytase was given when the Bm-N cells or fifth-instar silkworm larvae infected with a recombinant BmNPV containing the beta -galactosidase or phytase reporter gene driven by the polyhedrin promoter. We deduced that CTAB appeared to affect the virus bi-phasic life cycle stages and production pathways, resulting in an enhancement in BV production and a suppression of occluded virus (OV) production and expression of foreign genes controlled by the polyhedrin promoter.

摘要

一项实验研究旨在量化十六烷基三乙基溴化铵(CTAB)对家蚕核型多角体病毒(BmNPV)复制和 BmNPV ie-1 启动子转录活性的影响。结果表明,在感染野生型 BmNPV 的 Bm-N 细胞的培养基中添加 CTAB 至 0.1 µg ml(-1),可使出芽病毒(BV)滴度提高约 3.7 倍。在非感染昆虫细胞中,通过瞬时表达系统,在 0.1 µg ml(-1)CTAB 的存在下,BmNPV ie-1 启动子驱动的荧光素酶的瞬时表达水平提高了 3 倍以上。与 BV 滴度的升高相反,在 CTAB 浓度范围从 0 到 0.25 µg ml(-1)时,感染细胞内核中的多角体从 4.0×10(6)ml(-1)线性下降至 2.1×10(6)ml(-1)。当 Bm-N 细胞或感染含有β-半乳糖苷酶或植酸酶报告基因的重组 BmNPV 的五龄幼虫时,β-半乳糖苷酶或植酸酶的表达水平也呈现出相同的趋势,该报告基因由多角体蛋白启动子驱动。我们推断 CTAB 似乎影响病毒的双相生命周期阶段和产生途径,导致 BV 产量增加,而封闭病毒(OV)产量和多角体蛋白启动子控制的外源基因的表达受到抑制。

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