Chen Wen-Shuo, Chang Yen-Chung, Chen Ying-Ju, Chen Yu-Jie, Teng Chao-Yi, Wang Chung-Hsiung, Wu Tzong-Yuan
Institute of Molecular Medicine, National Tsing Hua University, Hsinchu, Taiwan.
J Virol Methods. 2009 Aug;159(2):152-9. doi: 10.1016/j.jviromet.2009.03.019. Epub 2009 Mar 26.
Recombinant baculoviruses are suitable for the high-level production of large multi-protein complexes. A tri-cistronic expression vector was constructed by the inclusion of two internal ribosome entry sites (IRESs). In this novel polycistronic vector, one single polyhedrin promoter controlled the transcription of a tri-cistronic transcript. Also, the first cistron was translated through a cap-dependent mechanism, while the second and third cistrons were translated by the IRESs derived from the 5' UTR of Rhopalosiphum padi virus (RhPV) and Perina nuda virus (PnV), respectively. The ratio of tri-cistronic gene expression levels produced by the three translational initiation modules is about 2:1:1 (cap:PnV IRES:RhPV IRES). This study indicates that polycistronic genes can be co-expressed at the translational level as in prokaryotic expression system by baculovirus biotechnology.
重组杆状病毒适用于大量多蛋白复合物的高水平生产。通过包含两个内部核糖体进入位点（IRES）构建了一个三顺反子表达载体。在这个新型多顺反子载体中，一个单一的多角体蛋白启动子控制三顺反子转录本的转录。此外，第一个顺反子通过帽依赖性机制进行翻译，而第二个和第三个顺反子分别通过源自麦长管蚜病毒（RhPV）和柿绒蚧病毒（PnV）5'UTR的IRES进行翻译。由三个翻译起始模块产生的三顺反子基因表达水平的比例约为2:1:1（帽：PnV IRES：RhPV IRES）。这项研究表明，多顺反子基因可以通过杆状病毒生物技术在翻译水平上像在原核表达系统中一样共表达。
