| 摘要: |
| 对青蛤（Cyclina sinensis）转录组文库筛选并克隆得到青蛤TRAF6（CsTRAF6）基因的cDNA序列，其开放阅读框为2337bp，编码778个氨基酸，分子量为86.59kDa。CsTRAF6蛋白包含RING型锌指结构，两个TRAF型锌指结构，coiled-coil结构域以及MATH结构域。经生物信息学及分子系统学分析表明CsTRAF6属于TRAF6家族的一员。利用实时荧光定量PCR方法检测了CsTRAF6基因在青蛤各个组织以及在Poly I：C侵染下在血淋巴中的时序性表达情况。结果表明，CsTRAF6基因在青蛤血淋巴、闭壳肌、鳃、性腺、外套膜和肝脏中普遍表达，其中在血淋巴中的表达水平最高。在Poly I：C刺激后的3h，青蛤血淋巴中CsTRAF6的表达量迅速升高，在6h达到最大值，说明青蛤的CsTRAF6基因参与了病毒类似物侵染下的免疫应答反应。 |
| 关键词: 青蛤 CsTRAF6 Poly I:C 荧光定量PCR |
| DOI：10.11693/hyhz20160600125 |
| 分类号: |
| 基金项目:天津市科委应用基础与前沿技术重点与青年项目资助，12JCZDJC22800号，14JCZDJC34200号，14JCQNJC14600号；天津市高等学校科技发展基金资助项目，20090608号；天津师范大学引进人才基金资助项目，5RL104号。 |
附件 |
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| CLONING OF CSTRAF6 GENE FROM CYCLINA SINENSIS AND IMMUNE RESPONSE UNDER POLY I: C STIMULATION |
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GAO Wei-Wei1, PAN Bao-Ping2, YAN Chun-Cai2
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1.Hexi District of Tianjin Community College For Staff, Tianjin 300203, China;2.College of Life Science, Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University, Tianjin 300387, China
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| Abstract: |
| The immune system of shellfish is non-specific and prevention against pathogens depends on hemolymph. The immune system can recognize the invading pathogenic microorganisms by Toll like receptors (TLRs), and then trigger the downstream signal transduction pathways. The tumor necrosis factor receptor associated factor 6 (necrosis factor receptor-associated factor tumor 6, TRAF6) gene plays an important role in the TLRs signaling pathway. We screened the cDNA sequence of TRAF6 (CsTRAF6) gene and cloned it from transcriptome library of clam Cyclina sinensis. The cDNA possessed an ORF of 2337bp encoding a protein of 778 amino acids; the predicted molecular weight was 86.59kDa and theoretical isoelectric point 6.16. The CsTRAF6 protein contains a RING-type zinc finger structure, two TRAF-type zinc finger structures, a coiled-coil domain, and a MATH domain. BLAST (the basic local alignment search tool) comparison and phylogenetic analysis show that CsTRAF6 is a member of the TRAF6 family. In addition, we detected the CsTRAF6 gene and their sequential expression in in all tissues by real-time PCR after Poly I:C infection. We found that the CsTRAF6 gene was generally expressed in hemolymph, adductor muscle, gill, gonad, mantle and liver, of which in hemolymph was the highest. The expression of CsTRAF6 in hemolymph injected with Poly I:C increased significantly in 3h and peaked at 6h, indicating that the gene was involved in the immune response against Poly I:C infection in C. sinensis. |
| Key words: Cyclina sinensis CsTRAF6 Poly I:C real-time PCR |
