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草鱼 g 型和 c 型溶菌酶的鉴定和表达分析。

Identification and expression analysis of the g-type and c-type lysozymes in grass carp Ctenopharyngodon idellus.

机构信息

Pearl River Fisheries Research Institute, Key Laboratory of Tropical & Subtropical Fish Breeding & Cultivation, Chinese Academy of Fishery Sciences, No. 1 Xing Yu Road, Guangzhou, PR China.

出版信息

Dev Comp Immunol. 2010 May;34(5):501-9. doi: 10.1016/j.dci.2009.12.009. Epub 2009 Dec 27.

DOI:10.1016/j.dci.2009.12.009
PMID:20034515
Abstract

Lysozyme is an important molecule of innate immune system for the defense against bacterial infections. We identified two genes encoding g-type and c-type lysozymes from grass carp Ctenopharyngodon idellus by the RACE method. The deduced amino acids of both lysozymes possessed typical structural residues and conserved catalytic sites similar to their counterparts across the species. In contrast, there was only 8.6% similarity of amino acid sequence between these two lysozymes. Phylogenetic analyses revealed that these two genes evolved at different rate. C-type lysozyme of grass carp was diverged early in the evolutionary history. Moreover, the expression patterns of these two genes differed. The mRNA levels of both genes were increased after bacterial infection, but the up-regulation of g-type lysozyme was much stronger than that of c-type lysozyme. We also showed that the c-type and g-type recombinant lysozymes possessed different lytic activities against fish bacterial pathogens. These results confirmed that both lysozymes play important roles in the defense of grass carp against bacterial infections. The g-type lysozyme may be induced for the defense against bacterial infections, while c-type lysozyme might be the main molecule for the house-keeping defense under normal conditions. These two types of lysozymes likely use different mechanisms to regulate their expressions.

摘要

溶菌酶是先天免疫系统抵抗细菌感染的重要分子。我们通过 RACE 方法从草鱼(Ctenopharyngodon idellus)中鉴定出两种编码 g 型和 c 型溶菌酶的基因。两种溶菌酶的推导氨基酸都具有典型的结构残基和保守的催化位点,与种间的对应物相似。然而,这两种溶菌酶的氨基酸序列只有 8.6%的相似性。系统发育分析表明,这两个基因以不同的速度进化。草鱼的 c 型溶菌酶在进化史上很早就分化了。此外,这两个基因的表达模式也不同。两种基因的 mRNA 水平在细菌感染后都增加了,但 g 型溶菌酶的上调幅度远大于 c 型溶菌酶。我们还表明,c 型和 g 型重组溶菌酶对鱼类细菌病原体具有不同的裂解活性。这些结果证实,两种溶菌酶在草鱼抵抗细菌感染的防御中都发挥了重要作用。g 型溶菌酶可能是为了抵抗细菌感染而被诱导产生的,而 c 型溶菌酶可能是正常情况下维持防御的主要分子。这两种类型的溶菌酶可能使用不同的机制来调节它们的表达。

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