青蛤(Cyclina sinensis)溶菌酶基因在鳗弧菌(Vibrio anguillarum)刺激下的表达  

EXPRESSION OF LYSOZYME GENE IN VIBRIO ANGUILLARUM-CHALLENGED CYCLINA SINENSIS

作　　者：潘宝平[1] 宋欣[1] 罗凯娅[1] 葛端阳[1] 高玮玮[1] 

机构地区：[1]天津师范大学生命科学学院细胞遗传与分子调控天津市重点实验室,天津300387

出　　处：《海洋与湖沼》2010年第6期901-906,共6页Oceanologia Et Limnologia Sinica

基　　金：天津市科委应用基础与前沿技术重点项目资助;09JCZDJC19300号

摘　　要：利用构建的SMART cDNA文库和高通量测序方法,得到了青蛤溶菌酶相关基因的全长,采用荧光定量RT-PCR方法分析了c型溶菌酶基因在鳗弧菌刺激下的表达变化。结果表明,青蛤有c型溶菌酶和i型溶菌酶基因,分别编码155和182个氨基酸,c型溶菌酶信号肽为21个氨基酸并具有典型的LYZ1结构域,i型溶菌酶信号肽为19个氨基酸,其结构域为Destabilase domain结构,用于识别和切断谷氨酰胺-甲酰胺与赖氨酸ε-氨基之间的肽键。荧光定量PCR结果显示,在鳗弧菌刺激后6—24h,青蛤血细胞中c型溶菌酶的表达量出现明显上调的趋势,与对照组有显著性差异(P<0.05),说明c型溶菌酶基因在青蛤的免疫应答中起重要的作用。Two lysozymes of Cyclina sinensis were cloned with high-throughput sequencing method to create the clams C. sinensis cDNA library construction. Real-Time PCR （RT-PCR） technique was applied in this study to produce the expression of c-type lysozyme by Vibrio anguillarum stimulated. The results showed that the full length cDNA of c-type and i-type lysozyme consisted of 804bp and 823bp respectively. The open reading frame （ORF） of c-type lysozyme encoded 155 amino acids （aa） including a signal peptide of 21aa at the N-terminus and a typical c-type lysozyme domain which was LYZ1 domain. The i-type lysozyme encoded 182 amino acids including a signal peptide of 19aa at the N-terminus and a Destabilase domain which cleaved isopeptide bonds formed by transglutaminase （Factor XIIIa） between glutamine gamma-carboxamide and the epsilon-amino group of lysine. Quantitative reverse transcriptase Real-Time PCR showed that the expression of c-type lysozyme gene increased after V. anguillarum infection from 6h to 24h. This trend was significant different from the control group （P0.05）. Our research showed that c-type lysozyme played an important role in the clam antibacterial immune.

关 键 词：青蛤 c型溶菌酶 i型溶菌酶 基因表达 

分 类 号：Q789[生物学—分子生物学]