| 摘要: |
| 本研究克隆得到了一个大竹蛏(Solen grandis)翻译控制肿瘤蛋白(TCTP)基因(SgTCTP)的cDNA 全长, 其序列全长为1055bp, 5′和3′端的非编码区(UTR)分别为54 和461bp, 开放阅读框(ORF)540bp, 编码179个氨基酸, 理论等电点为4.50, 预测分子量大小19.96kDa。通过荧光定量PCR法检测了SgTCTP在健康大竹蛏各组织中和病原相关分子模式(PAMPs)刺激后的表达规律, 结果表明: SgTCTP在检测组织外套膜、鳃、性腺、血细胞、肌肉和肝胰腺中都有表达, 其中在肝胰腺中的表达量最高。脂多糖(LPS)、肽聚糖(PGN)和葡聚糖(β-1,3-glucan)刺激都能诱导SgTCTP的表达量上调, SgTCTP 的表达量分别在LPS和PGN刺激后6和3h达到最高, 为空白对照的3.64和3.36倍; β-1,3-glucan 刺激后SgTCTP的表达量上升幅度最大, 在12h达到最高, 为空白对照的11.76 倍。SgTCTP可能作为急性时相蛋白参与大竹蛏的免疫应答 |
| 关键词: 大竹蛏 翻译控制肿瘤蛋白 免疫应答 荧光定量PCR |
| DOI：10.11693/hyhz201303007007 |
| 分类号: |
| 基金项目:山东省农业良种工程课题“优质高产抗逆贝类良种选育”, 2009—2013; 国家自然科学基金项目资助, 31202025 号; 水生动物营养与饲料“泰山学者”岗位经费资助 |
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| MOLECULAR CLONING AND EXPRESSION ANALYSIS OF A TRANSLATIONALLY CONTROLLED TUMOR PROTEIN (TCTP) GENE FROM SOLEN GRANDIS |
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WEI Xiu-Mei1, YANG Ding-Long1, YANG Jian-Min1, YANG Jia-Long2, XU Jie1, LIU Xiang-Quan1, ZHANG Yan-Min3
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1.Shandong Provincial Key Laboratory of Restoration for Marine Ecology, Shandong Marine Fishers Research Institute;2.Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences;3.Shandong Business Institute
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| Abstract: |
| In the present study, a translationally controlled tumor protein (TCTP) gene was identified from Solen grandis(designated as SgTCTP). The full-length cDNA of SgTCTP was of 1055bp, containing a 5’ untranslated region (UTR) of 54bp, a 3’ UTR of 461bp with a poly (A) tail, and an open reading frame (ORF) of 537bp encoding a polypeptide of 179 amino acids with the predicted molecular weight of 19.96kDa. The expression patterns both in tissues and towards pathogen associated molecular patterns (PAMPs) stimulation were then characterized by real-time PCR. SgTCTP was constitutively expressed in all tested tissues, including mantle, gill, gonad, hemocyte, muscle, and hepatopancreas, and it was highly expressed in hepatopancreas. The mRNA expression of SgTCTP could be induced by stimulation of β-1,3-glucan, LPS and PGN. The expression level of SgTCTP reached the peak at 12h and 3h post LPS and PGN stimulation, respectively. After β-1,3-glucan stimulation, SgTCTP expression reached the maximal level at 12h post stimulation, which was 11.76-fold compared with the blank group. All these results indicated that SgTCTP was an acute-phase protein involved in the immune response of S. grandis. |
| Key words: Solen grandis TCTP Immune response Real-time PCR |
