墨吉明对虾c型溶菌酶基因的克隆及表达  

Cloning and Expression of Gene of C-type Lysozyme from Fenneropenaeus merguiensis

作　　者：史黎黎[1] 周婷婷[1] 李逸群[1] 曾坤辉 陈兆明[1] 王成桂[1] 章双[1] SHI Li-li;ZHOU Ting-ting;LI Yi-qun;ZENG Kun-hui;CHAN Siuming;WANG Cheng-gui;ZHANG Shuang(Fisheries College, Guangdong Ocean University, Zhanjiang 524088, China)

机构地区：[1]广东海洋大学水产学院,广东湛江524088

出　　处：《广东海洋大学学报》2017年第3期8-13,共6页Journal of Guangdong Ocean University

基　　金：广东省自然科学基金-博士启动项目(2016A030310334);广东省自然科学基金-自由申请项目(2016A030313757);广东海洋大学优秀青年骨干教师特别资助计划项目(HDYQ2017003;HDYQ2015004)

摘　　要：克隆并鉴定墨吉明对虾(Fenneropenaeus merguiensis)的一种c型溶菌酶基因(Fm-Lyz),其cDNA全长为770 bp,包括71 bp的5′非编码区(UTR)、222 bp的3′UTR和477 bp完整开放阅读框,编码158个氨基酸。SMART分析显示,Fm-Lyz氨基酸序列包含1个LYZ1结构域和1个含18个氨基酸的信号肽。同源性分析发现,Fm-Lyz与斑节对虾(Penaeus monodon)溶菌酶同源性最高;进化树结果显示,Fm-Lyz与不同对虾的c型溶菌酶聚为一类。实时荧光定量PCR结果显示,Fm-Lyz在所测组织中均有表达,其中在血细胞中表达量最高。副溶血弧菌(Vibrio parahaemolyticus)和溶壁微球菌(Micrococcus lysodeikticus)感染后,Fm-Lyz表达量比对照组明显上调(P<0.05),最高表达量时间分别为感染后24、12 h,暗示Fm-Lyz参与了墨吉明对虾的抗菌免疫防御。The gene of a c-type lysozyme,namely Fm-lyz,from Fenneropenaeus merguiensis was cloned and identified.The full-length cDNA of Fm-Lyz is770bp with5'UTR of71bp,3'UTR of222bp and an open reading frame(ORF)of477bp,which encodes a putative protein of158amino acids including a LYZ1domain and a signal peptide with18amino acids.Multiple alignment analysis shows that Fm-Lyz shares the highest amino acid identity with Penaeus monodon.Phylogenetic analysis shows that Fm-Lyz is clustered into c-type lysozyme group of shrimp.qPCR analysis shows that Fm-Lyz is constitutively expressed in all the examined tissues with the highest expression in hemolymph.Upon Vibrio parahaemolyticus and Micrococcus lysodeikticus challenge,the expression of Fm-Lyz is significantly up-regulated compared with the control group(PBS group),and the time points of the highest expression is24h and12h,respectively,which suggests that Fm-Lyz may involve in anti-bacterial host defense.

关 键 词：墨吉明对虾 c型溶菌酶 基因 克隆 表达 

分 类 号：Q78[生物学—分子生物学] Q959.223.63