Abstract

To study the causes of impaired control of the activity of mobile genetic elements in the strains with the flamenco phenotype SS (w, flamenco mutant) and MS (w, flamenco mutant, active copy of gypsy), sequencing of these transcriptomes was performed. The D32 strain was used as control (laboratory wild type strain). An algorithm was developed for the search for amino acid substitutions in high-throughput RNA sequencing data using a triplet code for analysis. With the help of this algorithm, seven nonsense mutations were detected. The allele-specific PCR method confirmed the presence of five of the seven nonsense mutations found in silico. However, the detected nonsense mutations are not associated with the flamenco phenotype. A search for mutations in 89 genes of the RNA interference system in the SS and MS strains relative to the reference BDGP6 genome and the wild type D32 strain was carried out. No deletions, insertions, nonsense codons, and other disorders that can unambiguously lead to a change in the function of the gene are detected. To identify genes with specific expression for the strains with the flamenco phenotype, the transcriptomes of the SS and MS strains were compared with the control strains D-32, OregonR, and w1118. A set of 25 genes with differential expression was identified, among which two genes, sosie and CR45822, significantly changed the expression in the SS and MS strains. Both genes, directly or indirectly, are involved in oogenesis. Thus, the expression of the sosie and CR45822 genes can be used as a marker of the flamenco phenotype in the SS and MS strains.

中文翻译：

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果蝇逆转录转座子转位控制受阻的果蝇菌株的转录组分析

摘要

研究导致弗拉门戈表型SS（w，弗拉门戈突变体）和MS（w，弗拉门戈突变体，吉普赛人的有效拷贝）的菌株中移动遗传元件活性受到控制的原因），对这些转录组进行测序。将D32菌株用作对照（实验室野生型菌株）。开发了一种算法，该算法使用三联体代码进行分析以在高通量RNA测序数据中搜索氨基酸取代。借助该算法，检测到七个无意义的突变。等位基因特异性PCR方法确认了在计算机模拟中发现的七个无意义突变中的五个存在。但是，检测到的废话突变与弗拉门戈表型无关。相对于参考BDGP6基因组和野生型D32菌株，搜索了SS和MS菌株中RNA干扰系统的89个基因中的突变。没有检测到可以明确导致基因功能改变的缺失，插入，无意义密码子和其他疾病。为了鉴定具有弗拉门戈表型的菌株具有特异性表达的基因，将SS和MS菌株的转录组与对照菌株D-32，OregonR和w1118进行了比较。鉴定出25个具有差异表达的基因，其中两个基因sosie和CR45822显着改变了SS和MS菌株中的表达。两种基因都直接或间接参与卵子发生。因此，sosie和CR45822基因的表达可用作SS和MS菌株中弗拉门戈表型的标志。