Understanding_the_Molecular_Basis_of_Egg_Hatching_in_Trichuris_Species

My goal is to understand the molecular mechanisms behind the hatching process in T. muris. Our preliminary data has shown us that it is only possible to induce hatching at specific time points during the 8 week long embryonation process, namely weeks 7 and 8. We want to study the transcriptome and proteome of T. muris eggs arrested during embryonation. Using bulk RNA-seq to compare the expression profiles of the eggs at 6 weeks where hatching is not observed and in mature eggs; observing potential important changes in expression that explain the ability of the larvae to respond to bacteria. Secondly, our data has shown that the rate of hatching varies in the presence of different E. coli strains. Our in house strain of E coli displays two phase dependant phenotypes a “rough” type and “smooth” type, and the rough type is poor at inducing hatching. We aim to understand the genetic basis for this variation by generating a high quality reference genome of the two types. This reference genome will also allow us to to screen a high volume of E. coli mutants using the TraDIS approach; once we have generated transposon mutant libraries we will use T. muris eggs to perform a “pull down” style assay and sequence the pools of mutants that are either successful or unsuccessful at interacting with the egg. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/

本研究旨在解析T. muris（鼠鞭虫）孵化过程的分子机制。我们的预实验数据显示，仅在为期8周的胚胎发育进程中的特定时间点——即第7和第8周，方可诱导T. muris虫卵孵化。我们拟对胚胎发育停滞的T. muris虫卵开展转录组与蛋白质组学研究：通过批量RNA测序（bulk RNA-seq），对比未观察到孵化的第6周虫卵与成熟虫卵的表达谱，挖掘可解释幼虫对细菌应答能力的关键表达变化。其次，我们的数据表明，不同大肠杆菌（E. coli）菌株对T. muris的孵化率存在显著影响差异。本实验室自主培养的大肠杆菌菌株呈现两种两相依赖的表型：粗糙型（"rough"）与光滑型（"smooth"），其中粗糙型诱导虫卵孵化的能力较弱。我们计划通过构建这两种菌株的高质量参考基因组，解析该表型差异的遗传基础。该参考基因组还将支持我们利用TraDIS技术对大量大肠杆菌突变体进行筛选：在构建转座子突变体文库后，我们将使用T. muris虫卵开展"下拉"式结合实验，对能够与虫卵成功互作或无法结合的突变体池进行测序。本数据集属于预发布内容。如需了解威康信托桑格研究所（Wellcome Trust Sanger Institute）共享的预发布数据的规范使用方式（包括出版禁运期相关细则），请访问http://www.sanger.ac.uk/datasharing/