RNA干扰抑制因子P19对提高杆状病毒产量及Sf9细胞中转基因表达的功能分析

Functional analysis of RNAi suppressor P19 on improving baculovirus yield and transgene expression in Sf9 cells.

作者信息

Liu Yangkun, Zhang Lei, Zhang Yuanyuan, Liu Dandan, Du Enqi, Yang Zengqi

机构信息

College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, People's Republic of China.

China Institute of Veterinary Drug Control, Beijing, People's Republic of China.

出版信息

Biotechnol Lett. 2015 Nov;37(11):2159-66. doi: 10.1007/s10529-015-1910-5. Epub 2015 Jul 18.

DOI:10.1007/s10529-015-1910-5

PMID:26187316

Abstract

OBJECTIVE

To investigate whether RNA interference suppressor P19 derived from tombusvirus can enhance baculovirus yield and transgene expression.

RESULTS

A number of recombinant baculoviruses with P19 under the control of white spot syndrome virus immediate-early promoter, baculovirus early-to-late promoter, or P10 late promoter were constructed The budded virus titer and the expression levels of eGFP and luciferase were determined. P19 was clearly functional in Sf9 cells and could enhance baculovirus yield, eGFP and luciferase expression levels up to 6.8-, 1.8-, and 2.1-fold respectively, at 72 h post infection.

CONCLUSION

P19 enhanced baculovirus production and transgene expression, and thus has potential applications in baculovirus-based gene therapy and vaccine studies.

摘要

目的

研究番茄丛矮病毒来源的RNA干扰抑制因子P19是否能提高杆状病毒产量和转基因表达水平。

结果

构建了一系列重组杆状病毒，这些病毒中的P19分别受白斑综合征病毒立即早期启动子、杆状病毒早期至晚期启动子或P10晚期启动子的控制。测定了出芽病毒滴度以及eGFP和荧光素酶的表达水平。P19在Sf9细胞中具有明显功能，在感染后72小时，可分别将杆状病毒产量、eGFP和荧光素酶表达水平提高至6.8倍、1.8倍和2.1倍。