文章摘要
银鲳(Pampus argenteus) 3个营养代谢相关基因的克隆、表达及摄食水母对其表达模式的影响
Cloning and Expression of Three Genes of Pampus argenteus and the Effect of Jellyfish Feeding on Their Expression Patterns
投稿时间:2022-06-20  修订日期:2022-07-26
DOI:
中文关键词: 银鲳  营养代谢  基因  水母  表达模式
英文关键词: Pampus argenteus  nutrient metabolism  gene  jellyfish  expression pattern
基金项目:
作者单位邮编
练诗雅 中国水产科学研究院东海水产研究所
上海海洋大学水产与生命学院
上海海洋大学海洋学院 
200090
王亚冰 上海海洋大学水产与生命学院
中国水产科学研究院东海水产研究所 
王 倩 中国水产科学研究院东海水产研究所 
陈 润 中国水产科学研究院东海水产研究所 
岳彦峰 上海海洋大学水产与生命学院
中国水产科学研究院东海水产研究所 
苏志星 中国水产科学研究院东海水产研究所 
李云凯 上海海洋大学海洋学院 
彭士明 中国水产科学研究院东海水产研究所 200090
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中文摘要:
      本研究基于转录组学的分析结果,克隆分析了银鲳(Pampus argenteus)二肽酶(dehydropeptidase, dp1)、羧肽酶A(carboxypeptidases, cpa2l)和磺基转移酶(sulfotransferase, sult2)3个营养代谢相关基因,并探讨了其在消化吸收水母过程中发挥的功能。通过RACE技术克隆获得了银鲳dp1、cpa2l和sult2基因的cDNA全长序列。dp1基因全长2 522bp,包含一个1 272bp的开放阅读框(ORF),含有一个由23个氨基酸组成的信号蛋白肽和一个典型的酰胺水解酶超家族的结构域。cpa2l基因全长1 421bp,ORF长1 260bp,含有一个由16个氨基酸组成的信号蛋白肽和一个典型的M14金属羧肽酶家族的结构域。sult2基因全长1 834bp,ORF长714bp,含有一个典型的磺基转移酶家族的结构域。银鲳Dp1、Cpa2l和Sult2与蓝鳍金枪鱼(ThunnusSmaccoyii)同源性最高,系统进化树结果也显示银鲳dp1、cpa2l和sult2和蓝鳍金枪鱼对应基因聚在一个分支上,亲缘关系最近。运用Real-time PCR检测了dp1、cpa2l和sult2基因在不同组织中的表达水平及摄食水母对三个基因不同组织中表达规律的影响。结果发现银鲳dp1基因在肝脏中表达量最高(P<0.05),摄食水母组和未摄食水母组相比其在脑、鳃中表达量均极显著增加(P<0.01),在中肠和肾脏中显著增加(P<0.05),而在肌肉中显著下降(P<0.05)。cpa2l基因在未摄食水母组中中肠的表达量最高,而在摄食水母组中肾脏最高(P<0.05),摄食水母组和未摄食水母组相比其在肝脏中的表达量极显著增加(P<0.01),而在中肠和肌肉中却极显著下降(P<0.01)。sult2基因在肝脏中表达量最高(P<0.05),摄食水母组和未摄食水母组相比在中肠、脑、鳃、肝脏、肾脏中显著增加(P<0.05)。上述结果表明,dp1、cpa2l和sult2基因在银鲳摄食水母后机体营养物质的消化吸收和代谢过程中起重要的调节作用,基于摄食水母后3个基因组织表达模式的比较分析,推测dp1主要参与调控消化吸收和组织中营养素沉积的过程,cpa2l主要参与调控肝脏中营养代谢的过程,而sult2可能在整个消化吸收和代谢过程中均起到重要的调控作用。
英文摘要:
      In this study, three genes related to nutrition metabolism of Pampus argenteus, namely dehydropeptidase(dp), carboxypeptidase A(cpa) and sulfotransferase (sult), were cloned and analyzed based on transcriptome analysis results, and their functions in digestion and absorption of jellyfish were discussed. The full-length cDNA sequences of dp1,cpa2l and sult2 genes of Pampus argenteus were cloned by RACE technique. The total length of dp1 gene is 2 522bp, containing an open reading frame (ORF) of 1 272bp, a signal peptide composed of 23 amino acids and a domain of a typical amide hydrolase superfamily. The length of cpa2l gene is 1 421bp, and the length of ORF is 1 260bp. It has a signal peptide composed of 16 amino acids and a typical domain of M14 metal carboxypeptidase family. Sult2 gene was 1 834bp in length, ORF 714bp in length, and there was a typical domain of sulftase family. The amino acid sequences of dp1,cpa2l and sult2 of Pampus argenteus were the most homologous to bluefin tuna (Thunnus maccoyii). Phylogenetic tree results also showed that dp1,cpa2l and sult2 were closely related to the corresponding genes of Thunnus maccoyii. Real-time PCR was used to detect the expression levels of dp1, cpa2l and sult2 genes in different tissues and the effects of jellyfish ingestion on the expression levels of the three genes in different tissues. The results showed that the expression level of dp1 gene was the highest in liver (P<0.05). Compared with the non-feeding group, the expression levels of Pampus argenteus in brain and gill were significantly increased (P<0.01), and significantly increased in midgut and kidney (P<0.05), but significantly decreased in muscle (P<0.05). The expression level of cpa2l gene was the highest in the middle intestine of the non-eating jellyfish group, and the highest in the kidney of the eating jellyfish group (P<0.05). Compared with the non-feeding group, the expression level of Pampus argenteus was significantly increased in liver (P<0.01), but significantly decreased in midgut and muscle (P<0.01). The expression of sult2 gene was the highest in liver (P<0.05). Compared with the non-feeding group, the expression of Pampus argenteus in midgut, brain, gill, liver and kidney in the feeding group was significantly increased (P<0.05). These results suggest that dp1,cpa2l and sult2 genes play important roles in nutrient digestion, absorption and metabolism of Pampus argenteus after ingestion of jellyfish. Based on the comparative analysis of the tissue expression patterns of the three genes after ingestion of jellyfish, it is speculated that dp1 is mainly involved in the regulation of digestion, absorption and nutrient deposition in tissues, cpa2l is mainly involved in the regulation of nutrient metabolism in liver, and sult2 may play an important role in the whole process of digestion, absorption and metabolism.
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