Genetic Control of Organ Development in a Simple Eukaryote

本项目的主要目的是阐明一种名为FRT1的克隆基因诱导丛枝菌蘑菇(子实体)发育的分子机制。对从相应的cDNA和其他数据预测的蛋白质序列的分析表明，Ftr1编码一种膜相关蛋白，该蛋白通过某种形式的自我识别和非自我识别发挥作用，其活性受到交配型基因和相关遗传元件的活性的影响。这些概念将通过以下方法进行测试：1)确定FRT1在细胞内的位置的标签。2)利用体内诱变技术开始确定FRT1序列的哪些部分是活性所必需的。3)分离和鉴定连锁的DNA序列，该序列除了FRT1外，对于结实是必不可少的。这将使用DNA介导的转化方法来测试FRT1两侧的序列在适当的FRT1缺失突变中恢复结实表型的能力。4)通过使用FRT1和/或侧翼序列来分离和鉴定FRT1的替代等位基因，作为对公社链霉菌替代品系的克隆库的探针。在这个相对简单的真核生物中，通过研究这一重要发育过程的遗传调控而阐明的机械概念应该适用于理解更复杂、更难接近的生物体中的类似过程。

The primary objective of this project is to elucidate the molecular mechanisms by which a cloned gene called Frt1 induces development of mushrooms (fruiting bodies) in the woodrotting fungus Schizophyllum commune. Analysis of the protein sequence predicted from the corresponding cDNA, and other data, indicate that Ftr1 encodes a membrane- associated protein that functions through some form of self versus non-self recognition and that its activity is influenced by activity of the mating-type genes and a linked genetic element. These concepts will be tested by the following methods:1) a tag to determine the location of FRT1 within cells. 2) Employ the techniques of in vivo mutagenesis to start determining which parts of the Frt1 sequence are essential for activity. 3) Isolate and start characterizing the linked DNA sequence which, in addition to Frt1, is essential for fruiting this will be done using the method of DNA-mediated transformation to test the sequences flanking Frt1 for their ability to restore the fruiting phenotype in the appropriate Frt1 deletion mutations. 4) Isolate and start characterizing an alternate allele of Frt1 by using Frt1 , and/or flanking sequences, as probe to a clone bank of an alternate strain of S. commune. %%% Mechanistic concepts elucidated by studying genetic regulation of this important developmental process in this relatively simple eukaryote should be applicable to an understanding of similar processes in more complex, less accessible organisms.