Membrane fusion and infection abilities of baculovirus virions are preserved during freezing and thawing in the presence of trehalose
Membrane fusion and infection abilities of baculovirus virions are preserved during freezing and thawing in the presence of trehalose
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DOI：
10.1080/09168451.2019.1704396
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发表时间：
2019-12-21
影响因子：
1.6
通讯作者：
Tsumoto, Kanta
中科院分区：
文献类型：
--
作者：
Nakanishi, Kohei;Tomita, Masahiro;Tsumoto, Kanta
杆状病毒的芽状病毒（bv），如加利福尼亚自签名核多角体病毒（AcNPV），最近作为生物纳米材料进行了研究，并且需要一种不变质的长期储存方法。用天然存在的糖类如海藻糖作为低温保护剂对病毒粒子进行低温保存对于维持病毒的结构和功能是有用的。在这项研究中，我们研究了海藻糖在反复冻融循环中作为细菌性病毒冷冻保存的保护剂的作用：1)脂质体（多层囊泡，MLVs）和细菌性病毒之间的膜融合，2)表达rfp的细菌性病毒感染昆虫培养细胞（Sf9细胞），3)分别通过荧光脱冷实验、荧光显微镜和透射电子显微镜（TEM）研究了这些细菌性病毒的形态。结果表明，每次冻融循环后，BVs的质量都会恶化，而海藻糖的使用可以减少这种恶化，其程度与冰上储存的程度相似。
Budded viruses (BVs) of baculovirus such as Autographa californica nucleopolyhedrovirus (AcNPV) have recently been studied as biological nanomaterials, and methods for their longer-term storage without deterioration would be desirable. The cryopreservation of virions with a naturally occurring saccharide like trehalose as a cryoprotectant is known to be useful for maintaining the viral structure and function. In this study, we examined how useful trehalose is as protectant for BV cryopreservation during repeated freeze-thaw cycles: 1) membrane fusion between liposomes (multilamellar vesicles, MLVs) and BVs, 2) infection of insect culture cells (Sf9 cells) by RFP-expressing BVs, and 3) morphologies of these BVs were investigated by fluorescent dequenching assay, fluorescence microscopy, and transmission electron microscopy (TEM), respectively. The results suggest that the BVs deteriorate in quality with each freeze-thaw cycle, and this deterioration can be diminished with the use of trehalose to an extent similar to that seen with storage on ice.