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中国大鲵中鸡型和鹅型溶菌酶的表达及功能分析

Analysis on the expression and function of a chicken-type and goose-type lysozymes in Chinese giant salamanders Andrias davidianus.

作者信息

Yang Hui, Liu Ranran, Cui Dan, Liu Haixia, Xiong Dongmei, Liu Xiaolin, Wang Lixin

机构信息

College of Animal Science and Technology, Northwest A&F University, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100 China.

College of Animal Science and Technology, Northwest A&F University, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100 China.

出版信息

Dev Comp Immunol. 2017 Jul;72:69-78. doi: 10.1016/j.dci.2017.02.017. Epub 2017 Feb 24.

Abstract

Lysozymes as an important immune factor, play vital roles in innate immune response against pathogen infection. In the present study, one c-type and g-type lysozymes were identified from Chinese giant salamander (Andrias davidianus). They shared highly conserved structural features with lysozymes from other species. Spatial expression analysis revealed that AdlysC transcript was most abundant in liver and stomach, and least in muscle and brain. In contrast, the expression level of AdlysG was most abundant in liver and least in muscle and skin. The transcription level of c-type and g-type lysozymes were up-regulated after Aeromonas hydrophila infection in liver and spleen, indicating their participations in the immune response. Moreover, the recombinant AdlysC and AdlysG protein were produced and purified, and were used to investigate the lysozyme activity at different pH and temperatures. The optimal lytic activity was determined at pH 6.0 and at a temperature of 30 °C. Through the minimal inhibitory concentration test, the rAdlysC and rAdlysG exhibited apparent antibacterial activity against both Gram-positive and Gram-negative bacteria with a variable concentration. In conclusion, it is the first report of lysozymes in A. davidianus, and c-type and g-type lysozymes should be involved in the innate immune response of A. davidianus.

摘要

溶菌酶作为一种重要的免疫因子,在针对病原体感染的固有免疫反应中发挥着至关重要的作用。在本研究中,从中国大鲵(Andrias davidianus)中鉴定出一种c型和一种g型溶菌酶。它们与其他物种的溶菌酶具有高度保守的结构特征。空间表达分析显示,AdlysC转录本在肝脏和胃中最为丰富,在肌肉和大脑中最少。相比之下,AdlysG的表达水平在肝脏中最高,在肌肉和皮肤中最低。c型和g型溶菌酶的转录水平在嗜水气单胞菌感染后在肝脏和脾脏中上调,表明它们参与了免疫反应。此外,制备并纯化了重组AdlysC和AdlysG蛋白,并用于研究不同pH和温度下的溶菌酶活性。在pH 6.0和30°C的温度下确定了最佳裂解活性。通过最低抑菌浓度试验,rAdlysC和rAdlysG对革兰氏阳性菌和革兰氏阴性菌均表现出明显的抗菌活性,且浓度不同。总之,这是首次报道大鲵中的溶菌酶,c型和g型溶菌酶应参与大鲵的固有免疫反应。

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