Yokohama Research Center, JNC Co., Yokohama, Japan.
Photochem Photobiol. 2022 Nov;98(6):1293-1302. doi: 10.1111/php.13479. Epub 2021 Aug 25.
The genomic structure of the Cypridina luciferase gene in Vargula hilgendorfii (formerly Cypridina hilgendorfii) was determined with three λ phage clones (λ34, λ45, and λ61). The luciferase genes in clones λ34 and λ61 consisted of 13 exons and 12 introns, and clone λ45 only contained exons 1-5. The splicing sites of the luciferase genes in λ34 and λ61 were conserved completely with the consensus sequence. The translated luciferases had 555 amino acid residues, which were over 98.6% identical to those of cDNA clones as previously reported. In contrast, each intron in clones λ34, λ45, and λ61 varied significantly in length. To explain the variation of intron length among the three V. hilgendorfii luciferase genes, genomic DNA was isolated from a single V. hilgendorfii specimen, and the regions from exon 1-3 of the luciferase gene were amplified by polymerase chain reaction (PCR). PCR products with various lengths were detected and were confirmed as the luciferase gene fragments by Southern blot analysis. Furthermore, DNA sequence analysis indicated that at least seven luciferase gene groups might be present in the genome of a single specimen. Thus, multiple Cypridina luciferase genes exist in the genome of a single V. hilgendorfii specimen.
采用 3 个 λ噬菌体克隆（λ34、λ45 和 λ61）确定了海萤荧光素酶基因在海豆芽（以前称为 Cypridina hilgendorfii）中的基因组结构。克隆 λ34 和 λ61 中的荧光素酶基因由 13 个外显子和 12 个内含子组成，而克隆 λ45 仅包含外显子 1-5。λ34 和 λ61 中荧光素酶基因的剪接位点与一致序列完全保守。翻译的荧光素酶有 555 个氨基酸残基，与之前报道的 cDNA 克隆高度一致，超过 98.6%。相比之下，克隆 λ34、λ45 和 λ61 中的每个内含子在长度上差异显著。为了解释三种海豆芽荧光素酶基因中内含子长度的变化，从单个海豆芽标本中分离基因组 DNA，并通过聚合酶链反应（PCR）扩增荧光素酶基因的外显子 1-3 区域。检测到具有不同长度的 PCR 产物，并通过 Southern blot 分析确认为荧光素酶基因片段。此外，DNA 序列分析表明，单个标本的基因组中可能存在至少 7 个荧光素酶基因群。因此，单个海豆芽标本的基因组中存在多个海萤荧光素酶基因。
