DEVELOPING A PROTEIN EXPRESSION SYSTEM FOR SCORPION BETA TOXINS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 蝎子毒液是一种非凡的鸡尾酒，含有许多生物活性物质。许多不同的大分子都是用多肽来表示的，这是最好的表征。今年夏天的奖学金将为重新提交给NIH的一项更大的提案提供初步数据。我们更大的项目检查了毒素多肽的种内可变性。这个更大的项目侧重于一种基因组学方法，该方法表征了来自无毒类似物种的几个种群的毒素多肽，该物种在医学上具有重要的意义。这种方法将允许确定毒素多肽的可变性，并提供对毒素多肽中的氨基酸替代如何产生降低医疗效果的毒液的洞察。为了完成这项工作，我们需要一种强大的方法来生产蝎子毒素蛋白。因此，我们将把这个INBRE项目的重点放在亚克隆和毒素表达的目标上。该项目可导致以下方面的进一步工作：1)确定贝特钠毒素DNA和蛋白质序列中的关键核苷酸和氨基酸差异；2)确定无毒物种(Centruroides Vittatus)的贝特钠毒素的种内变异性是否也见于毒性较强的物种(Centruroides Exilicauda)的毒素变异性；3)对肽和蛋白质结构进行分子建模。该项目还可能提供细节，最终更好地阐明神经细胞的膜通道调节，并可能展示如何人工改变蝎子毒素多肽以达到医疗目的。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Scorpion venom is a remarkable cocktail that contains many bioactive agents. Many different macromolecules are represented with peptides as the best characterized. This summer fellowship will provide preliminary data for a larger proposal to be resubmitted to NIH. Our larger project examines intraspecific variability in toxin peptides. This larger project focuses upon a genomics approach that characterizes venom toxin peptides from several populations of a non toxic similar species to the medically important Centruroides sculpturatus. This approach will allow the determination of toxin peptide variability and provide insight into how amino acid substitutions in toxin peptides can produce venoms with reduced medical effects. To complete this work, we require a robust means of producing scorpion toxin proteins. Thus, we will focus this INBRE project on the objectives of subcloning and toxin peptide expression. This project can lead to further work to 1) identify key nucleotide and amino acid differences in sodium beta toxin DNA and protein sequence; 2) determine if intraspecific variability in sodium beta toxin from a non-toxic species (Centruroides vittatus) is also seen in toxin variability in a more toxic species (Centruroides exilicauda); 3) conduct molecular modeling of peptide and protein structures. This project may also provide details that eventually better illustrate membrane channel regulation in neurons and may show how scorpion toxin peptides can be artificially altered for medicinal purposes.