Ministry of Agriculture, Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Silkworm Biotechnology, Zhenjiang, Jiangsu, 212018, China.
Cytotechnology. 2003 Jan;41(1):37-44. doi: 10.1023/A:1024231023015.
To characterize the effects of cetyltriethylammonium bromide (CTAB) on the transcription of gp64 promoter from Bombyx mori nucleopolyhedrovirus (BmNPV), the plasmid pBmgp64Luc used in transient expression assay system was constructed by using the luciferase gene as a reporter under the control of BmNPV gp64 promoter. When the Bombyx mori cells (Bm-N) were transfected with the pBmgp64Luc, different treatments were undertaken. We found that the transient expression activity of luciferase could not be augmented directly by CTAB treatment alone, but could be enhanced more than 2 times by BmNPV treatment alone at a multiplicity of infection (MOI) of 0.5. Through co-treatment with 0.1 microg ml(-1) of CTAB and BmNPV at a MOI of 0.5, the enzymatic activity increased 5.21 times. We presumed that the stimulation of transcription of BmNPV gp64 promoter by CTAB was mediated by viral factors from BmNPV. In addition, the time curves of luciferase activity in cells transfected with pBmgp64Luc and transactivated by virus were observed.
为了研究十六烷基三乙基溴化铵（CTAB）对家蚕核型多角体病毒（BmNPV）gp64 启动子转录的影响，构建了瞬时表达检测系统用的质粒 pBmgp64Luc，该质粒中报告基因为荧光素酶，受 BmNPV gp64 启动子调控。将 pBmgp64Luc 转染家蚕细胞（Bm-N）后，进行了不同处理。我们发现 CTAB 单独处理不能直接增强荧光素酶的瞬时表达活性，但单独用 BmNPV 处理在感染复数（MOI）为 0.5 时可使活性增强 2 倍以上。通过用 0.1 μg/ml CTAB 和 MOI 为 0.5 的 BmNPV 共同处理，酶活性增加了 5.21 倍。我们推测 CTAB 对 BmNPV gp64 启动子转录的刺激是由 BmNPV 的病毒因子介导的。此外，还观察了转染 pBmgp64Luc 并被病毒反式激活的细胞中荧光素酶活性的时间曲线。
