DEVELOPING A PROTEIN EXPRESSION SYSTEM FOR SCORPION BETA TOXINS

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 蝎毒是一种了不起的鸡尾酒，含有许多生物活性剂。 许多不同的大分子以肽为代表，作为最好的表征。 这个夏季奖学金将为重新提交给NIH的更大提案提供初步数据。 我们更大的项目研究毒素肽的种内变异。 这个更大的项目集中在一个基因组学的方法，从几个种群的无毒的类似物种的特点毒液毒素肽的医学重要的雕刻Centruroides。 这种方法将允许确定毒素肽的变异性，并提供洞察毒素肽中的氨基酸取代如何产生具有降低的医疗效果的毒液。 为了完成这项工作，我们需要一种强大的生产蝎毒素蛋白的方法。 因此，我们将把这个INBRE项目的重点放在亚克隆和毒素肽表达的目标上。 该项目可以导致进一步的工作，以1）确定钠β毒素DNA和蛋白质序列中的关键核苷酸和氨基酸差异; 2）确定来自无毒物种（Centruroides vittatus）的钠β毒素的种内变异性是否也见于毒性更强物种（Centruroides exilicauda）的毒素变异性; 3）进行肽和蛋白质结构的分子建模。 该项目还可能提供细节，最终更好地说明神经元中的膜通道调节，并可能显示如何人为改变蝎毒素肽用于医学目的。

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.