单环刺螠泛素基因的克隆及表达  被引量：2

The clone and expression analysis of ubiquitin gene in the Urechis unicinctus

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作　　者：张立涛 崔巍 陈茜 毕愿坤 ZHANG Litao;CUI Wei;CHEN Qia;BI Yuankun(College of Biological Sciences,Jining Medical University,Rizhao 276826,China)

机构地区：[1]济宁医学院生物科学学院,日照276826

出　　处：《济宁医学院学报》2018年第6期396-399,共4页Journal of Jining Medical University

基　　金：山东省基金联合专项(ZR2016CL17);济宁医学院博士基金(JY2015BS15)

摘　　要：目的以单环刺螠为研究对象,克隆其泛素基因并分析其器官表达规律,为其功能的进一步研究提供理论依据。方法采用BLAST方法在单环刺螠转录组中获得泛素基因序列,设计引物扩增其开放阅读框(open reading frame,ORF)并经测序对其进行验证,随后对序列进行多序列比对和进化分析,进一步确定序列的准确性,最后采用q RT-PCR方法对泛素基因在单环刺螠各器官的表达规律进行分析。结果克隆获得单环刺螠泛素基因,其ORF为234 bp,编码76个氨基酸,氨基酸序列与其他物种的泛素极其相似。单环刺螠泛素基因在体腔液中表达量最低,在后肠和中肠的表达量是体腔液表达量的2. 39倍和2. 78倍,体壁的表达量是体腔液表达量的3. 46倍,而肛门囊表达量达到体腔液表达量的4. 55倍。结论本研究成功克隆获得单环刺螠泛素基因,其在单环刺螠各器官的表达规律:体腔液<肠道<体壁<肛门囊。Objective Urechis unicinctus was chosen as the objective of study and then we cloned the ubiquitin gene and obtained its organ expression characteristics,which can offer the theoretical foundation for further research of ubiquitin function.Methods The sequence of the ubiquitin was obtained from the transcriptome of the U.unicinctus by BLAST.Afterwards,its ORF was amplified by the designed primers and verified by sequencing.For further validation,multiple sequence alignment and evolutionary analysis was conducted.Finally,qRT-PCR was used to obtained organ expression characteristics of ubiquitin gene.Results We cloned the ubiquitin gene with the ORF of 234 bp coding 76 amino acids from U.unicinctus.The ubiquitin sequence of U.unicinctus shared high similarity with other species.The ubiquitin expression was the lowest in thecoelomic fluid,followed by the midgut(2.39 hold)and hindgut(2.78 hold),the body wall(3.46 hold),and the highest in the anal sac(4.55 hold).Conclusion In the study,the ubiquitin gene was successfully cloned from U.unicinctus and its expression feature in different organs was displayed:coelomic fluid<intestine<body wall<anal sacs.

关 键 词：单环刺螠 泛素 表达规律 

分 类 号：Q178.53[生物学—水生生物学]

 

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