文献类型：期刊文献

英文题名：A lef5-deficient baculovirus expression system with no virion contamination and promoting secretion

作者：Hu, Die[1];Xie, Xueman[1];Zhang, Tong[1];Yu, Yue[1];Xu, Zhenhe[1];Zhang, Yuanxing[2];Liu, Qin[1,2,3]

机构：[1]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, 130 Meilong Rd, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Shanghai Frontiers Sci Ctr Optogenet Tech Cell Met, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2023

卷号：365

起止页码：20

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20230713598227);WOS:【SCI-EXPANDED(收录号:WOS:000942537400001)】；

基金：This work was supported by the Shanghai Frontiers Science Center of Optogenetic Techniques for Cell Metabolism grant 2021 Sci & Tech 03-28 (Shanghai Municipal Education Commission) and the Funda- mental Research Funds for the Central Universities (222201211716) .

语种：英文

外文关键词：Autographa californica multiple; nucleopolyhedrovirus; Baculovirus expression vector; Insect cell; Lef5; Virus particle

摘要：The baculovirus expression system (BEVS) is widely used in biomanufacturing. However, massive late and very late genes are expressed during baculovirus infection of the host cell, and the protein aggregation has a significant negative impact on the intercellular environment of cells and organelle function. Baculovirus particle contamination also hinders the purification of particulate products. In this study, a novel baculovirus vector of deficient-lef5 (Ac-Delta lef5) was established to prevent the production of baculovirus particles. It could transduce cells and replicate viral DNA but the expression of the late and very late genes was reduced so that no virus was packed and budded. The Ac-Delta lef5 expression system was successfully used to express a secreted protein, thyroid peroxidase (TPO). Compared to wild-type baculovirus (Ac-wt), Ac-Delta lef5 delayed the decline in cell viability and prolonged the harvest period from 4 to 6 days after infection. When expressed in the Ac-Delta lef5 system, 98% of TPO was secreted extracellularly, which was about 1.8 times that of the Ac-wt system. Meanwhile, the transcription levels of protein folding-related genes were significantly increased. The results show that the Ac-Delta lef5 system is a potential novel viral-free baculovirus expression system, which omits the virion removal operation in biomanufacturing.