| 摘要: |
| 利用构建的SMART cDNA文库和高通量测序方法, 得到了青蛤溶菌酶相关基因的全长, 采用荧光定量 RT-PCR 方法分析了 c 型溶菌酶基因在鳗弧菌刺激下的表达变化。结果表明, 青蛤有 c型溶菌酶和 i 型溶菌酶基因, 分别编码 155 和 182 个氨基酸, c 型溶菌酶信号肽为 21 个氨基酸并具有典型的 LYZ1结构域, i型溶菌酶信号肽为 19个氨基酸, 其结构域为 Destabilase domain结构, 用于识别和切断谷氨酰胺?-甲酰胺与赖氨酸ε-氨基之间的肽键。荧光定量 PCR 结果显示, 在鳗弧菌刺激后6—24h, 青蛤血细胞中 c 型溶菌酶的表达量出现明显上调的趋势, 与对照组有显著性差异(P<0.05), 说明 c 型溶菌酶基因在青蛤的免疫应答中起重要的作用。
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| 关键词: 青蛤, c 型溶菌酶, i 型溶菌酶, 基因表达 |
| DOI：10.11693/hyhz201006016016 |
| 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, 09JCZDJC19300 号 |
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| EXPRESSION OF LYSOZYME GENE IN VIBRIO ANGUILLARUM-CHALLENGED CYCLINA SINENSIS |
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PAN Bao-Ping, SONG Xin, LUO Kai-Ya, GE Duan-Yang, GAO Wei-Wei
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College of Life Sciences, Tianjin Key Laboratory of Cyto-Genetical and Molecular Regulation, Tianjin Normal University
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| Abstract: |
| Two lysozymes of Cyclina sinensis were cloned with high-throughput sequencing method to create the clams C. sinensis cDNA library construction. Real-Time PCR (RT-PCR) technique was applied in this study to produce the expression of c-type lysozyme by Vibrio anguillarum stimulated. The results showed that the full length cDNA of c-type and i-type lysozyme consisted of 804bp and 823bp respectively. The open reading frame (ORF) of c-type lysozyme encoded 155 amino acids (aa) including a signal peptide of 21aa at the N-terminus and a typical c-type lysozyme domain which was LYZ1 domain. The i-type lysozyme encoded 182 amino acids including a signal peptide of 19aa at the N-terminus and a Destabilase domain which cleaved isopeptide bonds formed by transglutaminase (Factor XIIIa) between glutamine gamma-carboxamide and the epsilon-amino group of lysine. Quantitative reverse transcriptase Real-Time PCR showed that the expression of c-type lysozyme gene increased after V. anguillarum infection from 6h to 24h. This trend was significant different from the control group (P<0.05). Our research showed that c-type lysozyme played an important role in the clam antibacterial immune.
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| Key words: Cyclina sinensis, c-type lysozyme, i-type lysozyme, Gene expression |
