厚壳贻贝抗菌肽的分子多样性分析【毕业论文】.docVIP

本科毕业论文 （20 届） 厚壳贻贝抗菌肽的分子多样性分析 专业：生物科学 目录 摘要 I Abstract II 引言 1 1 材料与方法 2 1.1 动物与试剂 2 1.1.1 实验动物 2 1.1.2 试剂 2 1.2 实验方法 2 2 实验结果与分析 4 2.1 厚壳贻贝血淋巴抗菌肽的分离纯化 4 2.2 厚壳贻贝血淋巴抗菌肽的质谱鉴定 6 2.3 厚壳贻贝血淋巴的抑菌活性 9 2.4 部分厚壳贻贝血淋巴抗菌肽的鉴定 10 3 讨论 12 3.1 讨论 12 小结 15 参考文献 17 致谢 19  摘要 [摘要] 厚壳贻贝（Mytilus coruscus）具有重大经济价值[Abstract]：Mytilus coruscus is widely spreaden all through the littoral extent of East China Sea and of important economic value. Abundant peptides with antimicrobial activity were identified from hemocytes of mussel in previous works. To elucidate the compositive characterization of antimicrobial peptides from blood cell and serum of Mytilus coruscus hemocytes, high performance liquid chromatogram were used and several fractions with distinct antimicrobial activity were isolated from blood cell and serum of Mytilus coruscus, respectively. Mass spectrometry combined cDNA data analysis were performed to characterize the molecular mass and annotation of some peptides. Some of antimicrobial peptides identified in this work were considered belong to Mytilins or Myticins family with molecular mass ranged from 3000 to 7000 Da. The difference between blood cell and serum antimicrobial peptides mainly exists in molecular composition and antimicrobial spectrum. In addition, a novel peptide with strong antimicrobial activity was found in serum of Mytilus coruscus and the molecular mass of this peptide is more than 10000 Da. The results of this work build the foundation for further research on the molecular diversity of antimicrobial peptides, immune mechanisms of Mytilus coruscus, and screening for new peptides with applicable value. [Key words]：Mytilus coruscus; antimicrobial peptides; blood cell; serum; Mass spectrometry 引言 抗菌肽是，分子量通常分布于27000 Da左右，由2060个氨基酸残基组成低等生物由于缺乏特异性免疫系统，因而抗菌肽对于低等生物的免疫系统具有特别重要的意义，同时，抗菌肽也因其高效的抗菌活性而备受关注。抗菌肽是海洋生物抗菌肽研究中的内容，目前已从贻贝血细胞和血清中分离鉴定了种抗菌肽家族，分别为，，MGD和，主要分离自地中海贻贝（Mytilus galloprovincialis和紫贻贝Mytilus edulis）[3-7]。厚壳贻贝（Mytilus coruscus）采自浙江舟山海域饲养于恒温水族箱（℃）TF