Pak J Biol Sci. 2022 Jan;25(6):501-508. doi: 10.3923/pjbs.2022.501.508.
<b>Background and Objective:</b> In the puerulus phase, which is not pigmented, identification of lobster species based on morphological characteristics is still difficult identity, so it is necessary to identify based on molecular characters. This study aimed to analyze the mitochondrial subunit I cytochrome oxidase (COI) gene characters of the puerulus of lobster species. <b>Materials and Methods:</b> The data can be useful for developing lobster seed identification methods based on DNA characteristics. Location of lobster sampling in Staring Bay, coastal waters of Moramo District, South Konawe Regency, Indonesia. The molecular characterization method is carried out in several stages, namely specific primer design, DNA preparation, PCR with specific primers, DNA sequencing and DNA sequence analysis. Characteristics of COI gene fragments were analyzed using BLAST analysis, restriction enzyme analysis and phylogenetic tree analysis. <b>Results:</b> The results showed that DNA was successfully isolated with a high level of purity. The results of the amplification of the COI gene fragment showed thick and firm bands and formed a single band measuring 751 bp with 249 amino acids. Based on the BLAST analysis shows that the COI gene fragment has 99% similarity with <i>Panulirus homarus</i>. Based restriction enzyme analysis shows that the site of recognition and restriction enzyme cutting position in the <i>Panulirus</i> COI gene fragment is the same as <i>Panulirus homarus</i> in Genebank, the <i>Ase</i>I enzyme in position 392 and <i>Psi </i>I in positions 47 and 106. <b>Conclusion:</b> Based on phylogenetic tree analysis, the COI gene fragment is in one group with <i>Panulirus homarus</i> and has a bootstrap value of 100% which shows that the nucleotide sequence is stable. The three analyzes show that the DNA source organism is the same species as <i>Panulirus homarus</i>.
在幼体期,龙虾尚未着色,基于形态特征进行物种鉴定仍然较为困难,因此有必要基于分子特征进行鉴定。本研究旨在分析龙虾幼体的线粒体亚单位 I 细胞色素氧化酶(COI)基因特征。材料与方法:这些数据可用于开发基于 DNA 特征的龙虾苗种鉴定方法。在印度尼西亚南科纳威摄政区莫拉莫区沿海的斯达林湾进行龙虾采样。分子特征分析方法分几个阶段进行,包括设计特异性引物、DNA 提取、特异性引物 PCR、DNA 测序和 DNA 序列分析。COI 基因片段特征分析采用 BLAST 分析、限制性内切酶分析和系统发生树分析。结果:结果表明,成功分离出具有高纯度的 DNA。COI 基因片段扩增结果显示,出现厚实、坚固的条带,形成一条 751bp 的单带,包含 249 个氨基酸。基于 BLAST 分析,COI 基因片段与<i>Panulirus homarus</i>具有 99%的相似性。基于限制性内切酶分析,<i>Panulirus</i>COI 基因片段的识别位点和酶切位置与 Genebank 中的<i>Panulirus homarus</i>相同,酶切位置为 Ase I 酶在 392 位,Psi I 酶在 47 位和 106 位。结论:基于系统发生树分析,COI 基因片段与<i>Panulirus homarus</i>在同一组,具有 100%的自举值,表明核苷酸序列稳定。这三种分析表明,DNA 来源的生物体与<i>Panulirus homarus</i>相同。