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Multiple Cypridina Luciferase Genes in the Genome of Individual Ostracods, Vargula hilgendorfii (Cypridina hilgendorfii)
Photochemistry and Photobiology ( IF 2.6 ) Pub Date : 2021-06-28 , DOI: 10.1111/php.13479 Satoshi Inouye 1
Photochemistry and Photobiology ( IF 2.6 ) Pub Date : 2021-06-28 , DOI: 10.1111/php.13479 Satoshi Inouye 1
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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.
中文翻译：
介形类个体基因组中的多个 Cypridina 荧光素酶基因，Vargula hilgendorfii（Cypridina hilgendorfii）
使用三个 λ 噬菌体克隆（λ34、λ45 和 λ61）确定了Vargula hilgendorfii（以前称为Cypridina hilgendorfii ）中的 Cypridina 荧光素酶基因的基因组结构。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，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共有序列完全保守。翻译的荧光素酶有 555 个氨基酸残基，与先前报道的 cDNA 克隆的相同度超过 98.6%。相反，克隆 λ34、λ45 和 λ61 中的每个内含子在长度上都有显着差异。为了解释三个V. hilgendorfii荧光素酶基因之间内含子长度的变化，基因组 DNA 从单个V. hilgendorfii标本和荧光素酶基因外显子 1-3 的区域通过聚合酶链反应 (PCR) 进行了扩增。检测不同长度的PCR产物，经Southern blot分析证实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七个荧光素酶基因组。因此，多个 Cypridina 荧光素酶基因存在于单个V. hilgendorfii标本的基因组中。
更新日期：2021-06-28
中文翻译：
介形类个体基因组中的多个 Cypridina 荧光素酶基因，Vargula hilgendorfii（Cypridina hilgendorfii）
使用三个 λ 噬菌体克隆（λ34、λ45 和 λ61）确定了Vargula hilgendorfii（以前称为Cypridina hilgendorfii ）中的 Cypridina 荧光素酶基因的基因组结构。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，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共有序列完全保守。翻译的荧光素酶有 555 个氨基酸残基，与先前报道的 cDNA 克隆的相同度超过 98.6%。相反，克隆 λ34、λ45 和 λ61 中的每个内含子在长度上都有显着差异。为了解释三个V. hilgendorfii荧光素酶基因之间内含子长度的变化，基因组 DNA 从单个V. hilgendorfii标本和荧光素酶基因外显子 1-3 的区域通过聚合酶链反应 (PCR) 进行了扩增。检测不同长度的PCR产物，经Southern blot分析证实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七个荧光素酶基因组。因此，多个 Cypridina 荧光素酶基因存在于单个V. hilgendorfii标本的基因组中。
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