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| 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 |
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孙金秋1, 徐莞媛1, 马杭柯1, 高威1, 欧阳乐飞1, 高焕1,2,3,4, 阎斌伦1,2,3,4
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1.江苏省海洋大学 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 连云港 222005;2.江苏省海洋生物产业技术协同创新中心 连云港 222005;3.江苏省农业种质资源保护与利用平台 南京 210014;4.江苏省海洋资源开发研究院(连云港) 连云港 222005
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| 摘要: |
| 本研究根据脊尾白虾(Exopalaemon carinicauda)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全长为1855 bp，其中，开放阅读框为1407 bp，5端非编码区为39 bp，3端非编码区为409 bp，共编码468个氨基酸，预测蛋白质的分子质量为152.55 kDa，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%。荧光定量分析结果显示，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+胁迫结果显示，其在低浓度(0.0100、0.0175和0.021 mmol/L) Cd2+胁迫中的表达模式基本一致，呈先升高后下降再升高再下降的趋势；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。 |
| 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 |
| DOI：10.19663/j.issn2095-9869.20181229001 |
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| Cloning and expression analysis of serine hydroxyl methyltransferase (SHMT) genes from Exopalaemon carinicauda |
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SUN Jinqiu1, XU Wanyuan1, MA Hangke1, GAO Wei1, OUYANG Lefei1, GAO Huan1,2,3,4, YAN Binlun1,2,3,4
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1.Jiangsu Key Laboratory of Marine Bioresources and Environment, Jiangsu Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005;2.Co-Innovation Center of Jiangsu Marine Bio-Industry Technology, Lianyungang 222005;3.Jiangsu Provincial Infrastructure for Conservation and Utilization of Agricultural Germplasm, Nanjing 210014;4.Marine Resource Development institute of Jiangsu (Lianyungang), Lianyungang 222005
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| Abstract: |
| In this study, based on the transcriptome of Exopalaemon carinicauda under hunger stress, 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 E. carinicauda SHMT gene is most homologous (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 E. carinicauda. Transcript levels were 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 alternately first increasing and then decreasing. Under high concentration of Cd2+ stress, the expression level was extremely low; and not even detected in these tissues. This suggests that high concentration of Cd2+ stress can inhibit the expression of the SHMT gene. The specific mechanism needs further study. |
| Key words: Exopalaemon carinicauda Serine hydroxyl methyltransferase Gene cloning Tissue expression Cd2+ stress |