农杆菌遗传转化秀丽隐杆线虫的筛选条件及构建物。

IF 2.6 3区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
PLoS ONE Pub Date : 2025-05-27 eCollection Date: 2025-01-01 DOI:10.1371/journal.pone.0325060
Eleanor C Warren, Andre E X Brown, Karen S Sarkisyan
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引用次数: 0

摘要

操纵模式生物内的基因表达对于反向基因实验是重要的,虽然产生转基因秀丽隐杆线虫的技术是可用的,但它们对于创建单个系是优化的。以秀丽隐杆线虫为模型创建转基因动物文库的能力将使新型实验成为可能,并将加速动物生理学的研究。在这里,我们描述了一系列的结构,旨在建立一个高通量的方法,介导的基因转移从农杆菌秀丽隐杆线虫转化。我们证明秀丽隐杆线虫能够以农杆菌为唯一食物来源存活,并筛选农杆菌介导的转化条件。虽然我们不能实现从农杆菌到秀丽隐杆线虫的常规基因转移,但我们认为该技术在进一步优化后具有潜力。该方法的成功将使秀丽隐杆线虫的快速和高通量转化成为可能,为目前可用的方法提供了改进。在这里,我们提供了测试的优化条件的详细信息,以及供科学界使用的t -二进制结构的有用资源。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Screening conditions and constructs for attempted genetic transformation of C. elegans by Agrobacterium.

Manipulating gene expression within a model organism is important for reverse genetic experimentation, and while techniques to generate transgenic C. elegans are available, they are optimised for creating individual lines. The ability to create libraries of genetically modified animals using C. elegans as a model would make new types of experiments possible and would speed up studies of animal physiology. Here, we describe a range of constructs designed to establish a high-throughput method of C. elegans transformation mediated by gene transfer from Agrobacterium. We demonstrate that C. elegans are able to survive on Agrobacterium as a sole food source, and screen conditions for Agrobacterium-mediated transformation in this organism. While we do not achieve routine gene transfer from Agrobacterium to C. elegans, we suggest that this technique has potential following further optimization. The success of the approach would enable rapid and high-throughput transformation of C. elegans, providing an improvement on currently available methods. Here we provide details of optimization conditions tested, and a useful resource of T-binary constructs for use by the scientific community.

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来源期刊
PLoS ONE
PLoS ONE 生物-生物学
CiteScore
6.20
自引率
5.40%
发文量
14242
审稿时长
3.7 months
期刊介绍: PLOS ONE is an international, peer-reviewed, open-access, online publication. PLOS ONE welcomes reports on primary research from any scientific discipline. It provides: * Open-access—freely accessible online, authors retain copyright * Fast publication times * Peer review by expert, practicing researchers * Post-publication tools to indicate quality and impact * Community-based dialogue on articles * Worldwide media coverage
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