| 摘要: |
| 利用荧光定量PCR方法检测了青蛤(Cyclina sinensis)在鳗弧菌(Vibrio anguillarum)胁迫下组织蛋白酶L基因(CsCPL)在各组织的表达差别及其在血淋巴中的时序表达关系。结果表明, CsCPL基因在青蛤血淋巴、肝脏、外套膜、鳃和闭壳肌等组织中普遍表达, 其中以血淋巴表达水平最高。在鳗弧菌刺激后3—96h青蛤的血淋巴中CsCPL的表达量均出现明显上调, 其中6h达到最大值并且与对照组有极其显著性差异(P<0.01), 说明该基因在青蛤免疫反应中具有重要作用。文章还对CsCPL基因进行了原核表达, 其产物蛋白分子量约35kDa, 经纯化及复性后具有明显的抑菌作用。 |
| 关键词: 青蛤 组织蛋白酶L 实时定量PCR 抑菌实验 |
| DOI：10.11693/hyhz20140100009 |
| 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 |
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| ANALYSIS OF EXPRESSION AND RECOMBINANT PROTEINS ACTIVITIES OF CATHEPSIN L GENE FROM CYCLINA SINENSIS |
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LIU Shi-meng,ZHAO Ting,ZHANG Hao and PAN Bao-ping
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College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin,300387,College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin,300387,College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin,300387,College of Life Sciences,Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin,300387
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| Abstract: |
| Measure the expression of Cathepsin L gene (CsCPL) between organizations in Cyclina sinensis stimulated by Vibrio anguillarum and time-dependent expression in hemolymph with RT-PCR method. The results showed that, CsCPL gene expresses abundantly in hemolymph, liver, mantle, gills and adductor muscle and other tissues in C. sinensis, with the highest expression level in hemolymph. The expression of CsCPL in hemolymph appeared a large number of increasement in 3—96 hours after V. anguillarum stimulation, and at 6 hours reach to maximum which was very significant different (P<0.01) with the control group, indicating that CsCPL gene plays an important role in the immune response in C. sinensis. Experiments also carried on the CsCPL genetic prokaryotic expression, and got recombinant protein about 35 kDa molecular weight, which has an obvious bacteriostatic action after purification and renaturation. |
| Key words: Cyclina sinensis cathepsin L Realtime PCR bacteriostatic test |
