EAGER Collaborative Research: Developing Transformation Technologies for Porphyra

海洋红藻的基因组正在由联合基因组研究所进行测序。序列信息将帮助科学家解决许多基本问题，因为紫菜属。属于古老的真核生物谱系，是重要的人类食物（“海苔”），具有复杂的生活史，而且-甚至与其他潮间带生物相比-具有异常的耐应激代谢。对新的基因组数据进行基于计算机的分析将足以解决一些研究问题，但大多数研究（例如，紫菜对极端干燥或强光的耐受性的基础）将需要基于生物信息学分析的实验方法。本项目将开发紫菜稳定遗传转化的关键技术，使这一实验工作成为可能。研究人员将专注于转化P. umbilicalis的中性孢子，因为这些大量产生的孢子缺乏细胞壁。这使得通过电穿孔等方法转化细胞成为可能。基于JGI测序项目提供的信息，将使用紫菜基因的天然紫菜启动子，并合成密码子优化的报告基因。将用选择性抗生素处理孢子以回收转化的孢子，并将在孢子成熟为成虫时评估转化的稳定性。PI和博士后助理将在两个参与实验室工作，以开发转换技术。博士后助理将研究转化技术开发过程中的生殖途径或应激生理学。由于全面的博士后培训和参与紫菜NSF研究协调网络，S/他将做好充分准备，以使个人和合作的进展与紫菜模型系统。该项目将使科学界能够在紫菜的实验研究中使用全基因组测序项目的数据，从而在代谢，进化和发育生物学领域取得根本性进展。

The genome of the marine red alga Porphyra umbilicalis is being sequenced by the Joint Genome Institute. The sequence information will help scientists address many fundamental questions, because Porphyra spp. belong to an ancient eukaryotic lineage, are important human foods ("nori"), have complex life histories, and---even compared to other intertidal organisms--- possess an unusually stress-tolerant metabolism. Computer-based analyses of the new genomic data will be sufficient to address some research questions, but most studies (e.g., the basis of Porphyra's tolerance to extreme drying or high light) will require experimental approaches based upon bioinformatics analyses. This project will develop the essential technology of stable genetic transformation in Porphyra to make such experimental work possible. The investigators will focus on transforming neutral spores of P. umbilicalis because these abundantly-produced spores lack a cell wall. This should make it possible to transform the cell by electroporation, among other approaches. Native Porphyra promoters of Porphyra genes will be used, based on information provided by the JGI sequencing project, and codon-optimized reporter genes will be synthesized. Spores will be treated with selective antibiotics to recover transformed sporelings, and the stability of the transformation will be assessed as sporelings mature to adults. The PIs and a postdoctoral associate will work across both participating laboratories to develop transformation technologies. The postdoctoral associate will investigate reproductive pathways or stress physiology during the development of transformation techniques. S/he will be well-prepared to make individual and collaborative advances with the Porphyra model system due to the comprehensive postdoctoral training and participation in the Porphyra NSF Research Coordination Network. This project will allow the scientific community to use the data from the whole genome sequencing project in experimental research on Porphyra, leading to fundamental advances in the areas of metabolism, evolution, and developmental biology.