Agricultural Genetic Engineering Research Institute, Agricultural Research Center, Giza, 12619, Egypt.
Mol Biotechnol. 2023 Mar;65(3):433-440. doi: 10.1007/s12033-022-00537-6. Epub 2022 Aug 18.
One of the most important factor that affects the efficient using of baculoviruses as a biopesticide is their sensitivity to UV irradiation. In this study, a photolyase gene (phr) of 1.4 kbp DNA fragment was cloned and characterized from Spodoptera littoralis granulovirus, an Egyptian isolate (SpliGV-EG1). A sequence of 466 amino acid were deduced when the gene was completely sequenced with a predicted molecular mass of ~ 55 kDa. Transcriptional regulation analyses revealed that phr transcripts were detected early at 6-h post-infection (hpi) and remained detectable until 72 hpi, suggesting their transcriptional regulation from a putative early promoter motif. An approximately ~ 55 kDa protein fragment was expressed from phr-induced bacterial culture and detected by SDS-PAGE and western blotting. In addition, direct exposure to UV irradiation resulted in a twofold decrease in SpliGV-EG1 occlusion bodies activation compared with Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) occlusion bodies which decreased with about 129-fold after exposure to UV irradiation based on median lethal concentration value (LC50). The obtained results suggested that the presence of photolyase gene possibly alters the inactivation of SpliGV-EG1-occluded bodies by UV irradiation. These results support the role and application of the photolyase protein to improve the damaged DNA repair mechanism as well as resistance of SpliGV to UV light inactivation.
影响杆状病毒作为生物农药有效利用的最重要因素之一是它们对紫外线辐射的敏感性。在这项研究中,从埃及分离株(SpliGV-EG1)的斜纹夜蛾颗粒体病毒中克隆并鉴定了一个 1.4 kbp 的 DNA 片段的光裂合酶基因(phr)。该基因完全测序后,推导出一个 466 个氨基酸的序列,预测分子量约为 55 kDa。转录调控分析表明,phr 转录本在感染后 6 小时(hpi)即可检测到,并可检测到 72 hpi,表明它们的转录调控来自一个假定的早期启动子基序。phr 诱导的细菌培养物中表达了一个约 55 kDa 的蛋白片段,并通过 SDS-PAGE 和 Western blot 检测到。此外,与斜纹夜蛾核多角体病毒(SpliNPV)的多角体相比,直接暴露于紫外线辐射会导致 SpliGV-EG1 包埋体的激活减少两倍,基于半数致死浓度值(LC50),暴露于紫外线辐射后,SpliNPV 的多角体减少了约 129 倍。研究结果表明,光裂合酶基因的存在可能改变了紫外线对 SpliGV-EG1 包埋体的失活作用。这些结果支持光裂合酶蛋白的作用和应用,以改善受损 DNA 的修复机制以及 SpliGV 对紫外线失活的抗性。