Key Laboratory of Silkworm Biotechnology, Ministry of Agriculture, Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang, Jiangsu, China.
Cytotechnology. 2002 May;39(1):27-35. doi: 10.1023/A:1022496703875.
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.
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。