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首页> 外文期刊>Fish & Shellfish Immunology >Identification, cloning and expression analysis ofTI Identification, cloning and expression analysis of Catechol-O-methyltransferase (COMT) gene from shrimp, Penaeus monodon and its relevance to salinity stress
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Identification, cloning and expression analysis ofTI Identification, cloning and expression analysis of Catechol-O-methyltransferase (COMT) gene from shrimp, Penaeus monodon and its relevance to salinity stress

机译：TI的鉴定，克隆和表达分析虾，对虾对苯二酚-邻甲基转移酶（COMT）基因的鉴定，克隆和表达及其与盐分胁迫的关系

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摘要

O-methyltransferase (OMT), a protein present ubiquitously in wide range of organisms plays significant role in methylation of small macro molecules for various functional and regulatory purposes. In crustaceans, OMT has functional role in growth, reproduction, ovarian development and molting. In the present study, suppression subtractive hybridization (SSH) performed using gill tissues of low (3ppt) and high (55ppt) salinity stressed shrimp Penaeus monodon resulted in identification of differentially expressed genes involved in signal transduction pathways, metabolism, defense proteins, DNA repair and synthesis, apoptosis, cell cycle regulation along with unknown and hypothetical proteins. Catechol-O-methyltransferase (COMT) a type of OMT was identified by SSH as one of the differentially expressed genes of shrimp P. monodon subjected to low and high salinity stress. The full length cDNA of COMT was cloned from the gills of P. monodon which consisted an open reading frame of 666 bp, encoding 221 amino acids. The ORF revealed one each of N-glycosylation and O-glycosylation sites and nine phosphorylation sites. The deduced amino acid sequence of COMT exhibited high sequence identity (92%) with COMT class of protein from Fenneropenaeus chinensis. Real time PCR analysis of the shrimp samples exposed to low salinity conditions at 3ppt revealed significant increase in expression of COMT transcripts in the guts at 24 h, 48 h, 1 week and 2 weeks, gills at 24 h and in the muscle tissues at 48 h, with maximum expression of the COMT levels by 5 fold in guts (1 week), 1 fold in gills (24 h) and 1.5 fold in muscle (48 h) respectively. The increased expression level of COMT at different time intervals in different tissues suggests a possible role of this gene in salinity stress tolerance in shrimps under low salinity conditions
机译：O-甲基转移酶（OMT）是一种广泛存在于多种生物中的蛋白质，在各种功能和调节目的的小分子大分子甲基化中起着重要作用。在甲壳类动物中，OMT在生长，繁殖，卵巢发育和蜕皮中具有功能性作用。在本研究中，使用低盐度（3ppt）和高盐度（55ppt）胁迫虾对虾对虾对虾组织进行的抑制消减杂交（SSH）导致鉴定出与信号转导途径，代谢，防御蛋白，DNA修复有关的差异表达基因以及合成，凋亡，细胞周期调控以及未知和假设的蛋白质。 SSH将邻苯二酚-O-甲基转移酶（COMT）一种OMT鉴定为遭受低盐度和高盐度胁迫的斑节对虾的差异表达基因之一。从斑节对虾的ill中克隆出COMT的全长cDNA，其由一个开放阅读框组成，长为666 bp，编码221个氨基酸。 ORF揭示了N-糖基化和O-糖基化位点各一个和9个磷酸化位点。推导的COMT氨基酸序列与来自费氏钩端螺旋体的COMT类蛋白具有很高的序列同一性（92％）。对3ppt暴露于低盐度条件下的虾样品进行的实时PCR分析显示，在24小时，48小时，1周和2周时肠道，24小时的g和48小时的肌肉组织中COMT转录物的表达显着增加h，COMT水平的最大表达量是肠道（1周）的5倍、,（24 h）的1倍和肌肉（48 h）的1.5倍。在低盐度条件下，不同时间间隔的不同组织中COMT的表达水平增加，表明该基因可能在虾的盐胁迫耐受性中发挥作用

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