Patterns of inheritance within the ciliate species Chilodonella uncinata - a poss

描述（由申请人提供）：本提案旨在阐明单一纤毛虫物种的遗传模式，以解决三个特定的假设。这些假设集中在(1)体细胞（大核）基因组和(2)种系（微核）基因组含量的变化，以及(3)表观遗传学在性偶联后体细胞基因组生成过程中的可能作用。这些假设是基于两个分离株（一个来自美国，一个来自波兰）的初步数据得出的，这两个分离株是一种形态种，Chilodonella uncinata。虽然分离物含有相同的核糖体dna和共享一些相同的蛋白质编码序列，但它们也显示出复杂的遗传模式。基因的重复拷贝在大核中是不同遗传的，在接合和系间杂交改变了至少一个基因的遗传模式，肌动蛋白。该项目的结果数据将揭示纤毛虫的遗传模式，以及可能与其他真核生物相似甚至同源的表观遗传过程。

DESCRIPTION (provided by applicant): This proposal aims to elucidate patterns of inheritance within a single species of ciliates to address three specific hypotheses. These hypotheses focus on variation in the content of (1) somatic (macronuclear) genomes and (2) germline (micronuclear) genomes, and on (3) the possible role of epigenetics in shaping the generation of somatic genomes following sexual conjugation. These hypotheses are based on preliminary data from two isolates (one from the USA and one from Poland) of a single morphospecies, Chilodonella uncinata. Although the isolates contain identical ribosomal DNAs and share some identical protein coding sequences, they also show complex patterns of inheritance. Duplicated copies of genes are differentially inherited in macronuclei following conjugation and cross between lines alter patterns of inheritance of at least one gene, actin. The resulting data from this project will shed light on patterns of inheritance within the ciliate Chilodonella uncinata, and also on epigenetic processes that are likely analogous, and perhaps even homologous, to events in other eukaryotes. To address these hypotheses, the proposed work will combine generation of clone libraries, qPCR, variant-specific PCR, and fluorescent microscopy. These methods are either already in use in the PIs laboratory or, in the case of fluorescent microscopy, currently under development. Finally, the proposed work will enable the PI to continue to involve students (both undergraduate and graduate) in her research program. The PI has a demonstrated record in recruiting students from traditionally underrepresented groups, and will continue with these efforts as part of the proposed work. The proposed work on ciliates is relevant to public health in that resulting discoveries will yield insights into developmentally-regulated genome processing in eukaryotic cells (cells with nuclei). The type of genome processing to be studied in this proposal is analogous to changes in genomes in human immune cells and in some types of cancers. Studies of microbial eukaryotes, including ciliates, have already led to discovery of numerous genome features that have broad relevance to humans, including the presence of telomeres on the end of chromosomes and the existence of self-splicing RNA.