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; 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。 物种之间的基因流是一个自然过程。无处不在的土壤细菌农杆菌和曲霉天然会将其一些DNA转移到植物上，从而导致产生冠状胆囊和毛茸茸的根部疾病。 转移DNA的能力已被利用用于基因工程，现在是将基因传递给植物细胞的首选方法。 尽管使用了广泛使用，但有关基因转移方式的许多机械问题仍然存在。在A. tumefaciens中，Vire2蛋白通过涂覆转移的DNA并促进其对植物细胞核的摄取来起着重要作用。但是，这似乎不是一种普遍的机制，因为曲霉的某些菌株有效地转移了DNA，但缺乏Vire2。 取而代之的是，这些菌株含有另一种可以代替Vire2的蛋白质胆汁，从而为核吸收转移的DNA提供了另一种途径。 显然，进入植物细胞后，胆囊定位在核中，它与附着在转移的DNA和植物蛋白（胆囊相互作用蛋白； GIP）的细菌驾驶蛋白相互作用。假设胆囊充当“分子拖拉机”，以将DNA转移到细胞核中。为了测试这一想法，将使用多种实验方法来了解胆囊和GIP如何通过与VIRE2无关的途径促进基因转移。更广泛的影响。 这些研究将增加对农杆菌介导的基因转移的理解，并确定该极为重要的基因输送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的教育和培训，并在普渡大学的博士后副学士。 此外，将招募来自布鲁克林学院的少数族裔大学生参加普渡大学的夏季研究实习。 经过夏季的经验，学生将继续在他们的家庭机构进行相关的动手研究。