Yan Laiqing, Li Guangdong, Deng Shoulong, Wang Likai, Wang Yiwei, Shen Zixia, Yin Depeng, Ji Pengyun, Wang Bingyuan, Liu Guoshi
State Key Laboratory of Animal Biotech Breeding, Key Laboratory of Animal Genetics, Breeding and Reproduction of the Ministry of Agriculture, Frontiers Science Center for Molecular Design Breeding (MOE), College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
National Center of Technology Innovation for Animal Model, National Health Commission of China (NHC) Key Laboratory of Comparative Medicine, Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences and Comparative Medicine Center, Peking Union Medical College, Beijing 100021, China.
Int J Mol Sci. 2025 Jan 13;26(2):606. doi: 10.3390/ijms26020606.
Melatonin is synthesized in multiple tissues and organs of pigs, and existing studies have shown the presence of the melatonin-synthesizing enzyme ASMT protein. However, the genomic information for the gene has been lacking. The aim of this study was to locate the genomic information of the gene in pigs using comparative genomics analysis and then obtain the coding region information through molecular cloning. First, using the NCBI Genome Data Viewer, we found that in most animals, the gene is often located next to the gene, with both genes arranged in the same direction. Similarly, the gene is commonly adjacent to the gene, also in the same orientation. We also discovered that the gene is frequently adjacent to the ASMT gene and arranged in the opposite direction. Using the "three-point localization" principle, we inferred the position of the gene based on the coordinates of and in pigs. Our results revealed that on the pig X chromosome, a gene called LOC110258194 is located next to the and genes, and its arrangement aligns with the gene in other species. Additionally, Ensembl contains a gene, ENSSSCG00000032659, at the same position, with completely overlapping exons, though it is not annotated as . Further analysis using the TreeFam tool from EMBL-EBI and the CDD tool from NCBI revealed that LOC110258194 and ENSSSCG00000032659 do not contain the typical Maf domain of and, thus, should not be annotated as , but rather as the gene. Using a slow-down PCR method for high-GC content genes, we successfully cloned the full CDS region of the pig gene and identified a new transcript missing Exon 6 and Exon 7. This transcript was submitted to NCBI and assigned the GenBank accession number MW847601. Our results represent the first successful localization of the gene in pigs, the first cloning of the gene's coding region, and the first discovery of a new transcript of the pig gene.
褪黑素在猪的多个组织和器官中合成，现有研究已表明存在褪黑素合成酶ASMT蛋白。然而，该基因的基因组信息一直缺失。本研究的目的是通过比较基因组学分析来定位猪中该基因的基因组信息，然后通过分子克隆获得编码区信息。首先，使用NCBI基因组数据查看器，我们发现，在大多数动物中，该基因通常位于基因旁边，两个基因排列方向相同。同样，该基因通常与基因相邻，方向也相同。我们还发现该基因经常与ASMT基因相邻且排列方向相反。利用“三点定位”原则，我们根据猪中基因和基因的坐标推断出该基因的位置。我们的结果显示，在猪的X染色体上，一个名为LOC110258194的基因位于基因和基因旁边，其排列与其他物种中的基因一致。此外，Ensembl在同一位置包含一个基因ENSSSCG00000032659，外显子完全重叠，尽管它未被注释为。使用EMBL-EBI的TreeFam工具和NCBI的CDD工具进行进一步分析发现 LOC110258194 和 ENSSSCG00000032659 不包含该基因的典型Maf结构域，因此不应注释为，而应注释为基因。使用针对高GC含量基因的慢扩增PCR方法，我们成功克隆了猪基因的完整CDS区域，并鉴定出一个缺失外显子6和外显子7的新转录本。该转录本已提交给NCBI并被赋予GenBank登录号MW847601。我们的结果代表了猪中该基因首次成功定位、该基因编码区的首次克隆以及猪基因新转录本的首次发现。
