New Approaches to Sponge-Microbial Symbioses

这项建议旨在更好地了解海绵-细菌之间的联系，特别是 确定涉及哪些细菌，以及这些关联有多稳定。了解以下内容 据预测，共生体将为药理活性海绵的水产养殖或 产生菌的培养。这些方法将具有巨大的潜力来解决 具有药用活性的海洋天然产品的供应问题。我们打算使用分子技术 例如变性梯度凝胶电泳法(DGGE)用于监测海绵中的微生物群落 以及应用最先进的化学分析来确定相应的二次 代谢物图谱。此外，我们还建议采用扩散生长的创新技术 室内(DGC)培养以前“不可培养”的海绵细菌。DGC本质上是 填充均匀海绵组织和琼脂的半透膜。这会使细菌固定。 在聚合物膜内，同时将细菌暴露在“海绵化学”中。这些 这些技术将使我们能够检验：1)海绵次生代谢物的变化是否与 海绵相关微生物群落的变化；2)海绵相关微生物群落的变化；2)海绵相关微生物群落的变化 海绵细菌；以及3)共生海绵细菌或海绵本身产生新的、有趣的、 药理活性化合物。除了对海绵-细菌共生的新见解(例如； 海绵细菌通才还是专家？)，我们将创建一个潜在独特细菌菌株的文库 用新的DGC技术从海绵中分离出来。分离出的海绵细菌将被培养出来 提取物，并筛选可能的新的抗癌或抗感染代谢物。任何一种情况下的活动 筛选后将对活性次生代谢物进行分离和结构鉴定。 为了有效地治疗癌症或结核病(由多重耐药细菌引起)等疾病，它是 重要的是开发新药。尽管从海军陆战队中分离出了几个潜在的候选药物 对于生物体来说，进一步的发展往往受到供应问题的阻碍。我们的研究旨在发现新的 来自微生物来源的候选药物，同时开发新的培养技术来解决 供应问题

This proposal aims to gain a better understanding of sponge-bacterial associations and, in particular, to determine which bacteria are involved and how stable these associations are. Understanding such symbioses is predicted to yield new approaches for the aquaculture of pharmacologically-active sponges or the cultivation of the producing bacterial strains. Such methods would have enormous potential to solve the supply problem of pharmaceutically-active marine natural products. We intend to use molecular techniques such as Denaturing Gradient Gel Electrophoresis (DGGE) to monitor the microbial communities in sponges as well as to apply state-of-the-art chemical analysis in order to determine the corresponding secondary metabolite profiles. Furthermore, we propose to adopt the innovative technique of Diffusion Growth Chambers (DGCs) to the cultivation of previously "unculturable" sponge bacteria. DGCs are essentially semi-permeable membranes filled with homogenized sponge tissue and agar. This immobilizes bacteria inside the polymer membranes, while simultaneously exposing bacteria to the "sponge chemistry". These techniques will allow us to examine whether: 1) changes in sponge secondary metabolites correlate with changes in sponge-associated microbial communities; 2) DGCs allow culturing of previously "unculturable" sponge bacteria; and 3) symbiotic sponge bacteria or sponges themselves yield new, interesting, pharmacologically-active compounds. In addition to new insights on sponge-bacterial symbiosis (e.g. are ; sponge bacteria generalists or specialists?), we will create a library of potentially unique bacterial strains isolated from sponges with the new DGC technique. Isolated sponge bacteria will be grown out and extracted, and extracts screened for possible new anticancer or antiinfectant metabolites. Activity in either screen will be followed by the isolation and structure elucidation of the active secondary metabolites. To effectively treat diseases such as cancer or tuberculosis (caused by multi-drug-resistant bacteria), it is important to develop new drugs. Although several potential drug candidates have been isolated from marine organisms, further development is often hampered by a supply problem. Our research aims to identify new drug candidates from microbial sources and simultaneously develop new culturing techniques to address the supply problem