增强携带 ADP 基因的重组人腺病毒 41 型（HAdV-41）的生长。

Enhanced growth of recombinant human adenovirus type 41 (HAdV-41) carrying ADP gene.

机构信息

Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, GA 30602, USA.

出版信息

Virus Res. 2013 Sep;176(1-2):61-8. doi: 10.1016/j.virusres.2013.05.003. Epub 2013 Jun 11.

DOI:10.1016/j.virusres.2013.05.003

Abstract

Human adenovirus type 41 (HAdV-41) has the potential to be constructed as a gene transfer vector for oral vaccine or gene therapy targeting gastrointestinal tract. Block in release of progeny virus from host cell severely affects the yield during virus amplification. In this study, HAdV-5 adenovirus death protein (ADP) gene was used to replace the open reading frames (ORFs) of the HAdV-41 E3 region to construct a backbone plasmid pAdbone41ADP. Recombinant adenoviral plasmids harboring ADP and GFP genes (pAd41ADP-GFP) were generated. Plaques were formed and HAdV-41-ADP-GFP virus was rescued after transfecting pAd41ADP-GFP into the packaging cell line 293TE32. When amplified on 293TE32 cells, HAdV-41-ADP-GFP virus released to the culture medium was 10-50 times more than control virus HAdV-41-GFP, which did not carry ADP gene. The results demonstrated that incorporation of the ADP gene substantially increased the yield of recombinant HAdV-41 virus through enhancing spread of progeny virus among packaging cells.

摘要

人腺病毒 41 型（HAdV-41）具有构建为针对胃肠道的口服疫苗或基因治疗的基因转移载体的潜力。阻止从宿主细胞中释放出子代病毒严重影响病毒扩增过程中的产量。在这项研究中，使用人 5 型腺病毒腺病毒死亡蛋白（ADP）基因取代 HAdV-41 E3 区的开放阅读框（ORF），构建了骨架质粒 pAdbone41ADP。生成了携带 ADP 和 GFP 基因的重组腺病毒质粒（pAd41ADP-GFP）。在将 pAd41ADP-GFP 转染到包装细胞系 293TE32 后，形成了噬菌斑并拯救了 HAdV-41-ADP-GFP 病毒。当在 293TE32 细胞上扩增时，与不携带 ADP 基因的对照病毒 HAdV-41-GFP 相比，HAdV-41-ADP-GFP 病毒释放到培养基中的量增加了 10-50 倍。结果表明，通过增强子代病毒在包装细胞之间的传播，引入 ADP 基因可显著提高重组 HAdV-41 病毒的产量。