Biology Department, Winthrop University, Rock Hill, SC 29733, USA.
Biology Department, Northeastern Illinois University, Chicago, IL 60625, USA.
Genes (Basel). 2022 Feb 7;13(2):312. doi: 10.3390/genes13020312.
The identification of mutants through forward genetic screens is the backbone of genetics research, yet many mutants identified through these screens have yet to be mapped to the genome. This is especially true of mutants that have been identified as mutagen-sensitive (), but have not yet been mapped to their associated molecular locus. Our study addressed the need for additional gene identification by determining the locus and exploring the function of the -linked mutagen-sensitive gene using three available mutant alleles: , , and . After first confirming that all three alleles were sensitive to methyl methanesulfonate (MMS) using complementation analysis, we used deletion mapping to narrow the candidate genes for Through DNA sequencing, we were able to determine that is the uncharacterized gene which encodes the ortholog of the highly conserved DNA2 protein that is important for DNA replication and repair. We further used the sequence and structure of DNA2 to predict the impact of the allele mutations on the final gene product. Together, these results provide a tool for researchers to further investigate the role of DNA2 in DNA repair processes in
通过正向遗传学筛选鉴定突变体是遗传学研究的核心，但通过这些筛选鉴定的许多突变体尚未被映射到基因组上。对于那些被鉴定为诱变敏感（）但尚未被映射到其相关分子基因座的突变体来说尤其如此。我们的研究通过确定位置并探索使用三个可用突变等位基因：、和的诱变敏感基因的功能，解决了额外基因鉴定的需求。首先，我们通过互补分析证实了所有三个等位基因对甲基甲磺酸（MMS）敏感，然后使用缺失作图来缩小候选基因的范围。通过 DNA 测序，我们能够确定是未被描述的基因，它编码高度保守的 DNA2 蛋白的同源物，该蛋白对于 DNA 复制和修复很重要。我们进一步使用 DNA2 的序列和结构来预测等位基因突变对最终基因产物的影响。这些结果为研究人员提供了一种工具，可进一步研究 DNA2 在 DNA 修复过程中的作用。
