Xenopus Microarrays In Model Of developmental tox

发育毒性模型中的非洲爪蟾微阵列

• 批准号: 6535183
• 负责人: Perry J Blackshear
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6535183
• 关键词: Xenopus; Xenopus oocyte; biotechnology; environmental toxicology; messenger RNA; microarray technology; nonmammalian vertebrate embryology; nucleic acid sequence; polymerase chain reaction; sequence tagged sites; technology /technique development

We have generated a Xenopus laevis expressed sequence tag (EST) collection from unfertilized eggs. These large cells contain all the maternal mRNA necessary for the first several hours and first cell divisions in developing Xenopus, since new DNA synthesis does not begin in this organism until after the mid-blastula transition. The use of eggs in this collection avoids the contamination of isolated oocytes with follicular cells. We have sequenced more than 18,000 ESTs from a normalized library from eggs; this has recently been subtracted and a new round of sequencing the subtracted library has been completed. This EST collection comprises a large fraction of the total Xenopus ESTs in the database at present. We have used PCR products from a subset of these ESTs to build prototype microarrays, which will ultimately be used in studies of early Xenopus development, including toxicology, drug testing, oligonucleotide or antibody perturbation of development, adding genetic information to the FETAX test system already available, etc. We have also funded an SBIR contract to a small company to use these ESTs in the development of microarrays. Early testing of these microarrays is currently underway. Finally, in collaboration with the sequencing group at Washington University in St. Louis, we are using clustering programs to generate a Xenopus "virtual genome project" of expressed sequence contigs.

我们已经产生了一个来自未剥皮鸡蛋的xenopus laevis表达的序列标签（EST）收集。这些大细胞包含最初几个小时的所有母体mRNA，并且在发育中的爪蟾中含有第一个细胞分裂，因为直到中间芽过渡后，新的DNA合成才开始在该生物体中开始。在该系列中使用卵可避免与卵泡细胞污染分离的卵母细胞。我们已经从鸡蛋的标准化文库中测序了18,000多个EST；最近已经减去了这一点，并完成了新的排序库。目前，该EST集合包括数据库中总Xenopus est的很大一部分。我们已经使用了这些EST子集的PCR产品来构建原型微阵列，最终将用于早期Xenopus开发的研究，包括毒理学，药物测试，寡核苷酸或开发的抗体扰动，将遗传信息添加到胎儿测试系统中。目前正在进行这些微阵列的早期测试。最后，与圣路易斯华盛顿大学的测序小组合作，我们正在使用聚类计划来生成表达序列重叠群的Xenopus“虚拟基因组项目”。