Collaborative Research: Novel Proteins Required for Gene Transfer to Plants

合作研究：基因转移至植物所需的新蛋白质

• 批准号: 1049836
• 负责人: Stanton Gelvin
• 金额: $ 30万
• 依托单位: Purdue University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1049836&HistoricalAwards=false
• 关键词: Collaborative; Research; Novel; Proteins; Required

Intellectual Merit. Gene flow between species is a natural process. The ubiquitous soil bacteria Agrobacterium tumefaciens and A. rhizogenes naturally transfer some of their DNA to plants, leading to production of crown gall and hairy root diseases. This ability to transfer DNA has been exploited for genetic engineering and is now the preferred method for delivering genes to plant cells. Despite the widespread use, many mechanistic questions remain about how the gene transfer takes place. In A. tumefaciens, the VirE2 protein plays a major role by coating the transferred DNA and facilitating its uptake into the nucleus of the plant cell. However, this does not appear to be a universal mechanism, as some strains of A. rhizogenes transfer DNA efficiently, but lack VirE2. Instead, these strains contain another protein, GALLS, that can substitute for VirE2, thus providing an alternate route for nuclear uptake of transferred DNA. Apparently, upon entry into plant cells, GALLS localizes inside the nucleus where it interacts with a bacterial pilot protein attached to the transferred DNA and with a plant protein (GALLS-Interacting Protein; GIP). GALLS is hypothesized to act as a "molecular tractor" to pull transferred DNA into the nucleus. To test this idea, multiple experimental approaches will be used to understand how GALLS and GIP promote gene transfer via a VirE2-independent pathway. Broader Impacts. These studies will increase understanding of Agrobacterium-mediated gene transfer and identify additional unknown components of this extremely important gene delivery system. This project will promote education and training of a doctoral student at Oregon State University and a postdoctoral associate at Purdue University. In addition, minority undergraduates from Brooklyn College will be recruited to participate in summer research internships at Purdue. After their summer experiences, the students will continue to engage in related hands-on research at their home institution.

智力优点。 物种之间的基因流动是一个自然过程。无处不在的土壤细菌根癌农杆菌和发根农杆菌自然地将其部分 DNA 转移到植物中，导致冠瘿和毛状根病的产生。 这种转移 DNA 的能力已被用于基因工程，现在已成为将基因传递到植物细胞的首选方法。 尽管广泛使用，但关于基因转移如何发生的许多机制问题仍然存在。在根癌农杆菌中，VirE2 蛋白通过包被转移的 DNA 并促进其被植物细胞核吸收而发挥着重要作用。然而，这似乎不是一种通用机制，因为一些发根农杆菌菌株可以有效地转移 DNA，但缺乏 VirE2。 相反，这些菌株含有另一种蛋白质 GALLS，它可以替代 VirE2，从而为转移 DNA 的核摄取提供了替代途径。 显然，进入植物细胞后，GALLS 定位在细胞核内，与附着在转移 DNA 上的细菌先导蛋白和植物蛋白（GALLS 相互作用蛋白；GIP）相互作用。假设 GALLS 充当“分子牵引器”，将转移的 DNA 拉入细胞核。为了测试这个想法，将使用多种实验方法来了解 GALLS 和 GIP 如何通过 VirE2 独立途径促进基因转移。更广泛的影响。 这些研究将增进对农杆菌介导的基因转移的了解，并鉴定这一极其重要的基因传递系统的其他未知成分。该项目将促进俄勒冈州立大学博士生和普渡大学博士后的教育和培训。 此外，布鲁克林学院的少数族裔本科生将被招募参加普渡大学的暑期研究实习。 暑期体验结束后，学生们将继续在所在机构从事相关的实践研究。