文章摘要
脊尾白虾丝氨酸羟甲基转移酶基因的克隆及其表达特征分析*
Cloning and Expression Analysis of Serine Hydroxyl Methyltransferase (SHMT) Genes from Exopalamon carinicauda
投稿时间:2018-12-29  修订日期:2019-01-22
DOI:
中文关键词: 脊尾白虾  丝氨酸羟甲基转移酶基因  基因克隆  组织表达  Cd2+胁迫
英文关键词: Exopalamon carinicauda  Serine hydroxyl methyltransferase  Gene cloning  tissue expression  Cd2+ stress
基金项目:
作者单位E-mail
孙金秋 江苏省海洋生物资源与生态环境重点实验室/江苏省海洋生物技术重点实验室 1171840685@qq.com 
徐莞媛 江苏省海洋生物资源与生态环境重点实验室/江苏省海洋生物技术重点实验室  
马杭柯 江苏省海洋生物资源与生态环境重点实验室/江苏省海洋生物技术重点实验室  
高威 江苏省海洋生物资源与生态环境重点实验室/江苏省海洋生物技术重点实验室  
欧阳乐飞 江苏省海洋生物资源与生态环境重点实验室/江苏省海洋生物技术重点实验室  
高焕 江苏省海洋生物资源与生态环境重点实验室/江苏省海洋生物技术重点实验室  
阎斌伦 江苏省海洋生物资源与生态环境重点实验室/江苏省海洋生物技术重点实验室 yanbinlun1962@163.com 
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中文摘要:
      本研究根据脊尾白虾转录组序列,采用cDNA末端快速扩增技术克隆获得了脊尾白虾丝氨酸羟甲基转移酶基因(SHMT)。该基因cDNA全长1855bp,其中开放阅读框1407bp,5’端非编码区为39bp,3’端非编码区为409bp,共编码468个氨基酸,预测蛋白质的分子质量为152.55kDa,理论等电点为4.90。同源性分析显示,脊尾白虾SHMT基因与甲壳类动物真宽水蚤(Eurytemora affinis)同源性最高,为96%。荧光定量分析结果表明,SHMT基因在脊尾白虾眼柄、胃、肝胰腺、心脏、鳃、肠、肌肉、腹索神经、皮下脂肪以及卵巢中均有表达,其中卵巢表达量最高,心脏次之。不同浓度Cd2+胁迫结果表明,其在低浓度(0.0100 mmol/L、0.0175 mmol/L和0.021 mmol/L)Cd2+胁迫中的表达模式基本一致,表现出先升高后下降再升高再下降的一个趋势;而在高浓度(0.0278 mmol/L)Cd2+胁迫中,该基因表达量很低,甚至不表达,说明高浓度Cd2+胁迫可以抑制该基因的表达,具体机制有待进一步研究。
英文摘要:
      In this study,based on the transcriptome of Exopalamon carinicauda stressed under hungry , one SHMT genes were obtained by rapid amplification of cDNA ends (RACE) (GenBank accession No: MH013225). The full-length cDNA of the SHMT gene was 1855 nucleotides and contained an open reading frame (ORF) of 1407 bp, encoding a protein of 468 amino acid residues, with the predicted molecular weight of 152.55 kDa and theoretical isoelectric point of 4.90. Multiple sequence alignment and phylogenetic analysis indicated that Exopalamon carinicauda SHMT gene has the most homology (96%) with that of Eurytemora affinis. The expression pattern of SHMT gene in different tissues was analyzed by qRT-PCR. The results showed that SHMT gene was expressed in all tissues of Exopalamon carinicauda. Transcript levels were the high in the heart and ovary, and were significantly higher in the ovary than in the other tissues. The results of different concentrations of Cd2+ stress showed that the expression patterns of lower cadmium stress were basically the same, showing a trend of increasing first, then decreasing, then increasing and then decreasing. In high concentration of Cd2+ stress, the expression level was very low, even not founded in above tissues. This suggests that high concentration of Cd2+ stress can inhibit the expression of this gene; the specific mechanism needs further study.
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