Development of DNA Insertion-specific Markers Based on the Intron Region of Oplegnathus punctatus itih4b for Genetic Sex Identification

IF 2.6 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Yuting Ma, Yongshuang Xiao, Zhizhong Xiao, Jun Li
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Abstract

Spotted knifejaw (Oplegnathus punctatus) is a significant marine fish species that exhibits pronounced sexual dimorphism, with males generally exhibiting greater weight and growth rates than females. Therefore, the farming of O. punctatus with a high proportion of males is beneficial for improving the quality and efficiency of the O. punctatus aquaculture industry. Furthermore, the development of a rapid technique in sexing O. punctatus fry will facilitate the selection and breeding of superior male varieties of O. punctatus. In this study, genome-wide scanning, comparative genomics, and structural variation analysis methods were employed to identify and extract the homologous region of the inter-alpha-trypsin inhibitor heavy chain 4 (itih4b) gene on the X and Y chromosomes from the complete genome sequence of O. punctatus. This analysis revealed the presence of a large segment of DNA insertion markers on the Y chromosome in the region. Itih4b plays an important role in the mechanisms that regulate inflammatory and immune responses in multicellular organisms. The method described here involved the design of a pair of primers to amplify two bands of 532 bp and 333 bp in males (individuals with DNA insertion variants in the intron of the itih4b gene). In females (individuals without DNA insertion), only one band of 333 bp could be distinguished by agarose gel electrophoresis. This method shortened the time required to accurately characterize intronic DNA insertion variants and genetic sexes in O. punctatus, thereby improving detection efficiency. This study has significant value for the large-scale breeding of O. punctatus all-male seedlings and provides a reference point for the study of intron variation regulation and RNA shearing in the itih4b gene.

Abstract Image

Abstract Image

基于Oplegnathus punctatus itih4b内含子区的DNA插入特异性标记的开发,用于基因性别鉴定。
斑点叉尾鮰(Oplegnathus punctatus)是一种重要的海水鱼类,具有明显的性二型,雄鱼的体重和生长速度通常高于雌鱼。因此,养殖雄性比例高的刀鲚有利于提高刀鲚养殖业的质量和效率。此外,开发一种快速的标枪鱼鱼苗性别鉴定技术将有助于选育优良的雄性标枪鱼品种。本研究采用全基因组扫描、比较基因组学和结构变异分析方法,从O. punctatus的完整基因组序列中识别并提取了X和Y染色体上的α-胰蛋白酶间抑制物重链4(itih4b)基因的同源区。该分析表明,在该区域的 Y 染色体上存在一大段 DNA 插入标记。Itih4b在多细胞生物的炎症和免疫反应调节机制中发挥着重要作用。本文介绍的方法是设计一对引物,在雄性个体(ih4b 基因内含子中存在 DNA 插入变体的个体)中分别扩增出 532 bp 和 333 bp 的两个条带。而在雌性个体(无 DNA 插入的个体)中,琼脂糖凝胶电泳只能分辨出一条 333 bp 的条带。这种方法缩短了准确鉴定O. punctatus基因内含子DNA插入变体和基因性别所需的时间,从而提高了检测效率。该研究对大规模培育O. punctatus全雄性幼苗具有重要价值,并为研究itih4b基因的内含子变异调控和RNA剪切提供了参考。
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来源期刊
Marine Biotechnology
Marine Biotechnology 工程技术-海洋与淡水生物学
CiteScore
4.80
自引率
3.30%
发文量
95
审稿时长
2 months
期刊介绍: Marine Biotechnology welcomes high-quality research papers presenting novel data on the biotechnology of aquatic organisms. The journal publishes high quality papers in the areas of molecular biology, genomics, proteomics, cell biology, and biochemistry, and particularly encourages submissions of papers related to genome biology such as linkage mapping, large-scale gene discoveries, QTL analysis, physical mapping, and comparative and functional genome analysis. Papers on technological development and marine natural products should demonstrate innovation and novel applications.
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