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目的:研究甲壳类动物先天免疫系统抗菌肽分子的多样性以及抗菌相关的酶活抑制功能。方法:通过大肠杆菌重组表达载体的构建与重组蛋白的表达与纯化,我们对凡纳对虾SWD分子抗菌相关的蛋白酶活性抑制作用进行了研究。结果:Lv-SWD分子能够被大肠杆菌正确表达,其中,Lv-SWD分子对应的mRNA的开放阅读框为279 bp,重组Lv-SWD分子包括92个氨基酸残基和一个组氨酸标签,理论分子量为12.9 kD;分泌型蛋白酶活性抑制实验的结果显示重组Lv-SWD分子具有较好的蛋白酶活性抑制作用。结论:Lv-SWD分子是在凡纳对虾中发现的一类新的抗菌肽分子,它具有良好的蛋白酶活性抑制作用,可能在无脊椎动物的先天免疫系统中发挥着免疫防御分子的作用。
OBJECTIVE: To study the diversity of antimicrobial peptide molecules in the innate immune system of crustaceans and the antimicrobial activity of antimicrobial peptides. Methods: Through the construction of the recombinant expression vector of Escherichia coli and the expression and purification of the recombinant protein, we studied the antimicrobial activity of SWD molecules against Proteus vannamei. Results: The Lv-SWD molecule was correctly expressed in E. coli. The open reading frame of Lv-SWD molecule was 279 bp, and the recombinant Lv-SWD molecule consisted of 92 amino acid residues and a histidine tag. The theoretical molecular weight Was 12.9 kD. The result of the inhibition test of secreted protease activity showed that the recombinant Lv-SWD molecule had a good protease activity inhibition effect. CONCLUSION: The Lv-SWD molecule is a new class of antimicrobial peptides found in Litopenaeus vannamei. It has a good inhibition of protease activity and may play an immune defensive molecular role in the innate immune system of invertebrates.