N. crassa H3K27me3 mutant hunt sequencing

A strain of N. crassa bearing two, silenced antibiotic resistance markers (NCU05173::hph and NCU07152::nat-1) in H3K27 methylated regions of the genome were exposed to UV mutagenesis and mutants defective in H3K27me-mediated silencing were selected on antibiotic containing media. The resulting mutants, which were in a Oakridge genetic background, were crossed to a genetically polymorhpic Mauriceville strain to allow for SNP mapping of the causative mutation. Progeny of these crosses that were resistant to hygromycin and/or nourseothricin (and therefore contained the resistance marker and the causative mutation to allow for expression) were pooled and sequenced. This project contains sequencing reads for the mutant progeny as well as the parental strains that were used to subtract any SNPs that were present in the parents.

一株携带两个沉默型抗生素抗性标记（NCU05173::hph与NCU07152::nat-1）、且该标记定位于基因组H3K27甲基化区域的粗糙脉孢菌（Neurospora crassa, N. crassa），经紫外线诱变处理后，在含抗生素的培养基上筛选出H3K27me介导的沉默功能缺陷突变体。所得突变体均处于Oakridge品系的遗传背景中，将其与遗传多态性显著的Mauriceville菌株进行杂交，以通过单核苷酸多态性（Single Nucleotide Polymorphism, SNP）定位引发沉默缺陷的致病变异。将杂交后代中对潮霉素（hygromycin）和/或诺尔丝菌素（nourseothricin）具有抗性的子代混合并测序——此类抗性子代既保留了原始抗性标记，又因存在致病变异而恢复了抗性标记的表达。本项目涵盖突变子代与亲本菌株的测序读段，用于扣除亲本基因组中固有存在的单核苷酸多态性信号。