論文 - 詳細
| RRC ID | 69509 |
|---|---|
| 著者 | Fujii T, Kakino K, Fukumori H, Hino M, Lee JM, Kusakabe T, Banno Y. |
| タイトル | Non-molting dwarf (nm-d) as a mutant of Bombyx mori with a defect in purine synthesis. |
| ジャーナル | Insect Biochem Mol Biol |
| Abstract |
There are several known non-molting mutations of the silkworm, Bombyx mori, including non-molting dwarf (nm-d). Larvae with this mutation hatch normally and start eating leaves, but die before the completion of the first ecdysis. Genetic analysis of the nm-d mutation would contribute to the isolation of essential genes for the larval development of lepidopteran insects. To identify the causative gene of the nm-d locus, we conducted RNA-seq based rough mapping. Using two sets of RNA-seq data, one from a pooled sample of normal larvae, and one from a pooled sample of nm-d larvae, the nm-d locus was narrowed to a 500 kb region. Among the genes located in this region, a nm-d-specific exon loss was identified in the Bombyx homolog of the ATIC (5-aminoimidazole-4-carboxamide ribonucleotide transformylase/Inosine 5'-monophosphate cyclohydrolase) (BmATIC) gene, which catalyzes the final two steps of the de novo purine biosynthetic pathway in mammals. PCR and subsequent sequencing analysis revealed that a region containing exon 9 of the BmATIC gene is deleted in the nm-d larvae. A knockout allele of the BmATIC gene (BmATICKO), that was generated using the CRISPR/Cas9 system, revealed that first instar knockout larvae died while exhibiting the dark brown larval body that is a typical feature of mutants that lack uric acid in the integument. Lethal larvae resulted from crosses between +/BmATICKO moths. The uric acid content in the whole-body of the first instar was drastically reduced in the nm-d larvae compared to normal larvae. These results indicated that the BmATIC gene is responsible for the nm-d phenotype, and that nm-d larvae have a defect in purine biosynthesis, including uric acid. We also discuss the possibility that the BmATIC mRNA is maternally transmitted to eggs. Our results indicated that RNA-seq based mapping using pooled samples is a practical method for the identification of the causative genes of lethal mutations. |
| 巻・号 | 138 |
| ページ | 103636 |
| 公開日 | 2021-11-1 |
| DOI | 10.1016/j.ibmb.2021.103636 |
| PII | S0965-1748(21)00119-3 |
| PMID | 34478812 |
| MeSH | Animals Insect Proteins / genetics* Insect Proteins / metabolism Larva / genetics Larva / growth & development Larva / metabolism Moths / genetics Moths / growth & development Moths / metabolism* Mutation* Purines / biosynthesis* |
| IF | 3.827 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 各媒体での言及数の合計 | 0 |
| リソース情報 | |
| カイコ | a47 o45 p50 |