School of Life Sciences, The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of Ministry of Education, Shandong University, Jinan, Shandong, China.
Fish Shellfish Immunol. 2010 Dec;29(6):1066-72. doi: 10.1016/j.fsi.2010.08.023. Epub 2010 Sep 9.
Lysozyme is an important immune effector and is widely distributed in many organisms. In the present study, two novel invertebrate-type lysozymes (Pclysi1 and Pclysi2) were cloned from red swamp crayfish, Procambarus clarkii. Alignment analysis showed that these two genes were different in catalytic residues. Results of RT-PCR showed that these two genes share similar tissue distribution patterns, and both were upregulated after bacteria challenge. The mature recombinant proteins were expressed in Escherichia coli system and purified. Activity analysis revealed that rPclysi1 had no muramidase activity and isopeptidase activity, but had antimicrobial activity. Meanwhile, rPclysi2 was muramidase-deficient and had no antimicrobial activity, but possessed isopeptidase activity. Above data suggest that Pclysi1 and 2 may offer different functions in the crayfish immunity.
溶菌酶是一种重要的免疫效应因子，广泛分布于多种生物中。本研究从克氏原螯虾（Procambarus clarkii）中克隆到两种新型的无脊椎动物溶菌酶（Pclysi1 和 Pclysi2）。序列比对分析显示这两个基因在催化残基上存在差异。实时定量 PCR 结果表明这两个基因具有相似的组织分布模式，且在细菌刺激后均被上调表达。成熟的重组蛋白在大肠杆菌系统中表达并纯化。活性分析表明 rPclysi1 没有溶菌酶活性和肽基内肽酶活性，但具有抗菌活性。同时，rPclysi2 缺乏溶菌酶活性且无抗菌活性，但具有肽基内肽酶活性。以上数据表明 Pclysi1 和 2 可能在螯虾免疫中发挥不同的功能。
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