红笛鲷(Lutjanus sanguineus)C型凝集素基因的克隆与组织表达分析  

Cloning and Tissue Expression Analysis of C-type Lectin Gene from Humphead Snapper (Lutjanus sanguineus)

作　　者：蔡佳[1,2,3] 张雪利[1,2,3] 吴灶和[2,3] 简纪常[1,2,3] 鲁义善[1,2,3] 冯东岳[4] 焦茂兴[5] 

机构地区：[1]广东海洋大学水产学院,湛江524088 [2]广东省水产经济动物病原生物学及流行病学重点实验室,湛江524088 [3]广东省水产经济动物病害控制重点实验室,湛江524088 [4]农业部全国水产技术推广总站,北京100125 [5]宜春学院,宜春336000

出　　处：《生物技术通报》2015年第5期179-185,共7页Biotechnology Bulletin

基　　金：广东高校国际合作创新平台项目(2013gjhz0008);十二五国家科技支撑计划(2012BAD17B02);广东教育厅科技创新重点项目(2012CXZD0026)

摘　　要：根据已知的海水鱼类C型凝集素基因的核苷酸保守区序列设计一对简并引物,先后通过RT-PCR和RACE PCR法从红笛鲷脾脏中首次克隆获得红笛鲷C型凝集素(CTL)基因的c DNA全长(登录号:AGT37609)。该序列长828 bp,开放阅读框663 bp,编码220个氨基酸。氨基酸序列分析显示,红笛鲷CTL基因氨基酸序列与其他物种CTL的相似性在30%-68%之间。系统进化分析表明,红笛鲷C型凝集素与鳉鱼、条石鲷、斜带石斑鱼CTL蛋白亲缘关系最近,聚成一支。通过荧光定量PCR分析红笛鲷基因的组织差异表达,红笛鲷CTL基因在肝、脾以及皮肤中表达水平较高,其次是头肾、肾、胃、肠及肌肉,在心脏与脑中表达量较低。According to conservative region of known C-type lectin （ CTL ） gene, a pair of degenerate primers were designed, and full length cDNA sequence of C-type lectin gene was amplified by RT-PCR and RACE PCR from spleen of humphead snapper, Lutjanus sanguineus （ GenBank accession number： AGT37609 ） for the first time. The total cDNA sequence of the gene was 828 bp, consisting of a 663 bp open reading frame （ ORF ） and encoding 220 amino acids. The deduced amino acid sequence of humphead snapper CTL shared 30%-68% identities with other species CTLs. Phylogenetic analysis showed that humphead snapper CTL was clustered closely with mummiehog, rock bream, and orange-spotted grouper CTL. Moreover, the mRNA expression levels in different tissues were analyzed by real time quantitative PCR. The result showed that humphead snapper CTL gene expressed in all tested tissues with highest level in liver, spleen and skin, moderate level in head kidney, kidney, stomach, intestine and muscle, and lowest level in heart and brain.

关 键 词：红笛鲷 C型凝集素 基因克隆 组织表达分析 

分 类 号：Q78[生物学—分子生物学]