Preservation of Dry Cells by Trehalose

许多在极度脱水中存活的生物（包括， 例如，酵母细胞、种子、真菌孢子和某些显微镜下的 动物）含有大量的糖，特别是 二糖蔗糖或海藻糖。 以前的研究已经表明 亚细胞部分如不稳定的酶和膜可以 如果它们在这些糖的存在下干燥，则保持完整。 如果 它们被干燥而没有糖，这些分子组合是 完全且不可逆地降解。 这种现象已经被习惯了。 开发保存蛋白质、膜和脂质体的方法， 水的缺乏，导致了几项专利， 行业 目前的项目扩展了有关亚细胞组分的信息 完整的细胞。傅里叶变换红外光谱用于 研究干燥后完整酵母细胞的膜状态 以导致各种量的海藻糖积累的方式。 这些研究旨在检验海藻糖 稳定完整细胞膜的方式与 在隔离的膜中。 海藻糖依赖性相图将是 产生的细胞，不仅应该测试这一假设， 细节，但也将立即在酵母实际使用 工业在提高脱水存活率方面的作用。 一种手段将 试图将海藻糖以干燥状态引入细胞， 脱水后通常无法存活 选择的路线是使用 酵母中发现的海藻糖转运蛋白。 分子生物学 技术将用于克隆这种转运蛋白的基因。 这项研究的合作者已经开发出了突变体 缺乏这种基因。 目的是通过互补获得该基因 这些变种人。 如果成功，基因将被导入 其他细胞在后续研究中。

Many organisms that survive extreme dehydration (including, for example, yeast cells, seeds, fungal spores, and certain microscopic animals) contain large quantities of sugars, particularly the disaccharides sucrose or trehalose. Previous studies have shown that subcellular fractions such as labile enzymes and membranes can be preserved intact if they are dried in the presence of these sugars. If they are dried without the sugars these molecular assemblages are completely and irreversibly degraded. This phenomenon has been used to develop means for preserving proteins, membranes, and liposomes in the absence of water, leading to several patents that are in use in industry. The current project extends the information about subcellular fractions to intact cells. Fourier transform infrared spectroscopy is used to study the state of membranes in intact yeast cells that have been dried in ways that result in accumulation of various amounts of trehalose. These studies are designed to test the hypothesis that trehalose stabilizes membranes in intact cells in much the same ways as it does in isolated membranes. A trehalose-dependent phase diagram will be produced for the cells that should not only test this hypothesis in detail, but will also be of immediate practical use in the yeast industry in effecting increased survival of dehydration. A means will be sought to introduce trehalose into cells in the dry state that would not normally survive dehydration. The route chosen is to use a transporter for trehalose that is found in yeasts. Molecular biology techniques will be used to clone the gene for this transporter. Mutants have already been developed by collaborators in this research that lack this gene. The aim is to obtain the gene by complementation of these mutants. If successful, the gene will be introduced into other cells in subsequent research.