| 摘要: |
| 利用青蛤(Cyclina sinensis)转录组文库信息, 设计引物并克隆得到了青蛤TLRs信号通路中的关键信号分子IκB(CsIκB)基因的cDNA序列。该基因的开放阅读框为1134bp, 编码377个氨基酸, 分子量为41.79kDa, 其结构具备IκB家族基因的典型特征, 包括C端的6个锚蛋白重复序列(ankyrin repeat, ANK), N端的DS64GILS68降解序列以及C端的酪蛋白激酶Ⅱ磷酸化位点(S373DDES377)。经BLASTX比对及分子系统学分析证明其是IκB家族的一员。进一步利用实时荧光定量PCR技术检测了CsIκB基因在青蛤不同组织及其在鳗弧菌(Vibrio anguillarum)胁迫下在特定组织中的时序性表达情况。结果表明, 该基因在样品的血淋巴、肝脏、外套膜、闭壳肌、鳃和性腺6种组织中普遍表达, 其中以血淋巴中表达水平最高。在鳗弧菌刺激后3h CsIκB在青蛤血淋巴中表达量明显上调, 并于6h达到最大值, 说明该基因参与了青蛤非特异免疫应答反应。 |
| 关键词: 青蛤 CsIκB 鳗弧菌 TLRs信号通路 荧光定量PCR |
| DOI：10.11693/hyhz20150500147 |
| 分类号: |
| 基金项目:天津市科委应用基础与前沿技术重点项目资助,12JCZDJC22800号,09JCZDJC19300号,14JCZDJC34200号。 |
附件 |
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| CLONING OF IκB GENE FROM CYCLINA SINENSIS AND THE EXPRESSION ANALYSIS UNDER VIBRIO ANGUILLARUM STIMULATION |
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WEI Xing, ZHANG Hai-Jing, PAN Bao-Ping
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College of Life Sciences, Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin Normal University, Tianjin 300387, China
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| Abstract: |
| Using clam Cyclina sinensis transcriptome library information, we designed primers then cloned the cDNA sequence of C.sinensis IκB (CsIκB), a key signal molecule in the TLRs signaling pathway.The cDNA possessed an ORF of 1134 bp coding for a protein of 377 amino acids with the predicted molecular weight of 41.79 kDa.CsIκB protein has the typical structural features with IκB family members, including the C-terminal six ankyrin repeats (ankyrin repeat, ANK), N-terminal DS64GILS68 degradation motif and a C-terminal casein kinase II phosphate sites (S373DDES377).BLASTX and phylogenetic analysis revealed that CsIκB is a member of IκB family.Further by using real-time PCR technique to detect the mRNA expression of CsIκB in various tissues and its temporal expression in specific tissue challenged with Vibrio anguillarum.The results showed that CsIκB is generally expressed in all the examined tissues with the highest expression level in hemolymph.The expression of CsIκB in hemolymph injected with V.anguillarum increased significantly at 3h and peaked at 6h, indicating that CsIκB involved in the non-specific immune response. |
| Key words: Cyclina sinensis CsIκB Vibrio anguillarum TLRs signaling pathway real-time PCR |
