日本沼虾表皮蛋白基因的克隆及表皮组织差异性表达分析  

Cloning and differential expression analysis of cuticle protein genes in different cuticular tissues from Macrobrachium Nipponense

作　　者：苗泽龙 吕艳杰[1] 张俊芳 杨洪[1] 宁黔冀[1] 

机构地区：[1]河南师范大学生命科学学院,河南新乡453007

出　　处：《河南师范大学学报（自然科学版）》2018年第2期79-83,共5页Journal of Henan Normal University(Natural Science Edition)

基　　金：国家自然科学基金(30940008);河南省教育厅科学技术研究重点项目(14A240003);河南师范大学青年科学基金(5101049279089)

摘　　要：为探究日本沼虾表皮蛋白(Macrobrachium nipponense cuticle proteins,MnCPs)表达模式与表皮组织差异的关系,根据日本沼虾表皮组织转录组测序结果,从头胸甲中克隆到1个含几丁质结合-4(chitin_bind_4)结构域的表皮蛋白(cuticle proteins,CPs)基因,命名为MnCP-2,经BLAST检索,MnCP-2与远海梭子蟹(Portunus pelagicus,ABM54465.1)有50%的相似性.以健康幼虾(体长3.0~4.0cm)的头胸甲、尾扇、游泳足和步足4种厚度存在差异的表皮组织为材料,分别提取C期、D_(0-2)期、D_(3-4)期和A期的RNA,采用Real-time PCR技术,分析了MnCP-2在蜕皮周期不同阶段的相对表达量.结果显示,MnCP-2在头胸甲、尾扇和游泳足中表达水平的峰值出现在D_(3-4)期,而在步足中则出现在D_(0-2)期.提示MnCP-2编码的蛋白可能在新外表皮的形成中发挥重要作用,它在表皮组织不同部位的差异性表达可能是导致表皮结构差异的原因之一.In order to understand the relationship between expression patterns of Macrobrachium nipponense cuticle proteins(MnCPs)and differences in cuticular tissues,agene containing chitin_bind_4 conserved domain called MnCP-2 was first cloned in carapace from M.nipponense,according to the result derived from transcriptome sequecing.BLAST search shows that MnCP-2 has 50%similarity with CPs fromPortunus pelagicus.RNA was extracted from the stage C,D_(0-2),D_(3-4) and A of healthy juvenile M.nipponense(body length 3.0~4.0 cm)in the carapace,tail fan,pleopod and pereiopod which have different thicknesses.Real-time PCR technique was used to analyze the expression levels of MnCP-2.The result showed MnCP-2 mRNA expression level was highest during stage D_(3-4) in carapace,tail fan and pleopod,while during stage D_(0-2) in pereiopod(P<0.05).Consequently,it suggests that MnCP-2 likely have an important role in forming new exocuticle,and their differential expressions in different cuticular tissues is possible one of the reasons why cuticle has different structures.

关 键 词：日本沼虾 表皮蛋白 头胸甲 尾扇 游泳足 步足 基因克隆 表达 

分 类 号：S966.1[农业科学—水产养殖]