Characterization of Synaptonemal Complexes and RecombinationNodules From Lilium longiflorum

长花百合联会复合体和重组结节的表征

• 批准号: 8918613
• 负责人: Stephen Stack
• 金额: $ 25.85万
• 依托单位: Colorado State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-03-01 至 1994-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8918613&HistoricalAwards=false
• 关键词: Characterization; Synaptonemal; Complexes; RecombinationNodules; Lilium

Correlative evidence suggests that synaptonemal complexes (SCs) and recombination nodules (RNs) are involved in chromosome synapsis and genetic recombination. However, little is known about the constituents of SCs and RNS that might help elucidate their precise funtion(s). It is proposed to isolate SCs and RNs from the plant species Lilium longiflorum and to characterize the proteins from isolated SCs/RNs by gel electrophoresis. Isolated SCs/RNs will also be injected into mice to elicit antibodies. Hybridomas will be made and screened against isolated SCs/RNs by immunofluorescence. The monoclonal antibodies identified will then be used (1) to identify SC/RN proteins in Western blots, (2) to examine developmental and tissue-specific expression of the SC/RN proteins, (3) to determine the cross-reactivity of the antibodies to other plant and animal SCs and RNs, and (4) to determine the relationship between nodules observed during four of the substages of meiotic prophase I (leptotene, zygotene, pachytene, and diplotene). These antibodies later can be used for affinity chromatography and/or immunoprecipitation to isolate SC/RN proteins in large quantities to determine their functions and to identify the genes coding for them. This work has the potential to lead to a real break-through in our understanding of the function(s) of SCs and RNs.

相关证据表明联会复合体（SC）和重组结（RN）参与染色体联会和基因重组。然而，人们对 SC 和 RNS 的成分知之甚少，而这可能有助于阐明它们的精确功能。 建议从植物百合中分离 SC 和 RN，并通过凝胶电泳表征分离的 SC/RN 的蛋白质。 分离的 SC/RN 也将被注射到小鼠体内以引发抗体。 将制备杂交瘤并通过免疫荧光针对分离的 SC/RN 进行筛选。 然后，所鉴定的单克隆抗体将用于 (1) 在蛋白质印迹中鉴定 SC/RN 蛋白，(2) 检查 SC/RN 蛋白的发育和组织特异性表达，(3) 确定抗体与其他植物和动物 SC 和 RN 的交叉反应性，以及 (4) 确定在减数分裂前期 I 的四个亚阶段期间观察到的根瘤之间的关系 （细线期、合子期、粗线期和双线期）。 这些抗体随后可用于亲和层析和/或免疫沉淀，以大量分离 SC/RN 蛋白，以确定它们的功能并鉴定编码它们的基因。 这项工作有可能使我们对 SC 和 RN 功能的理解取得真正的突破。