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作 者:谢飞 于洋[2] 周海龙[1] 李富花[2] XIE Fei;YU Yang;ZHOU Hai-Long;LI Fu-Hua(School of Life Sciences,Hainan University,Haikou 570228,China;Key Laboratory of Experimental Marine Biology,Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China)
机构地区:[1]海南大学生命科学学院,海南海口570228 [2]中国科学院海洋研究所实验海洋生物学重点实验室,山东青岛266071
出 处:《海洋与湖沼》2023年第5期1454-1462,共9页Oceanologia Et Limnologia Sinica
基 金:国家重点研发计划,2018YFD0900303号,2018YFD0901301号;中国科学院战略性先导专项,XDA24030105号;海南省基础与应用基础研究计划(高层次人才项目),2019RC077号。
摘 要:低密度脂蛋白受体相关蛋白(LRP)是一类细胞表面蛋白,属于一种内吞性受体,其在调节脊椎动物血脂平衡中的作用已被广泛报道。前期在凡纳滨对虾弧菌抗性家系和敏感家系的比较转录组分析中发现一种低密度脂蛋白受体相关蛋白基因的转录表达存在明显差异,为明确该基因在对虾抗弧菌感染中的作用,我们对该基因进行了全长cDNA克隆、表达特征分析和功能的初步研究。该基因在凡纳滨对虾中存在两个转录本,分别命名为LvLRP1-1和LvLRP1-2。LvLRP1-1的cDNA序列全长为916bp,编码226个氨基酸;LvLRP1-2的cDNA全长为787bp,编码183个氨基酸。两个转录本的cDNA序列高度一致,唯一的差别在于LvLRP1-1的cDNA比LvLRP1-2多129 bp,多编码43个氨基酸。组织表达分析显示LvLRP1-1在对虾的眼柄、表皮、脑和心脏中高表达,而LvLRP1-2仅在眼柄和表皮中呈现高表达。在副溶血弧菌感染对虾后, LvLRP1-1和LvLRP1-2的转录表达变化趋势一致,均在感染后3h其转录水平明显升高,6h之后又恢复到正常水平。利用双链RNA技术对LvLRP1-1和LvLRP1-2同时干扰后对对虾进行弧菌感染,发现干扰后的对虾体内弧菌的载量明显提高,死亡率显著上升,说明其在对虾抵抗弧菌感染的过程中发挥着重要作用。Low-density lipoprotein receptor-related protein(LRP)is a cell surface protein and an endocytic and cell signaling receptor.Its role in regulating blood lipid balance in vertebrates has been well known.Previously,one LRP gene was reported to have a significant difference in expression between the resistant and susceptible families to Vibrio in Litopenaeus vannamei by comparative transcriptome analysis.We cloned the full-length cDNA of the gene and analyzed its expression characteristics,and carried out a preliminary study on its function in order to clarify the role of the gene in L.vannamei anti-vibrio infection.Results show that there were two transcripts of this gene in L.vannamei,named LvLRP1-1 and LvLRP1-2.The length of LvLRP1-1 is 916 bp encoding 226 amino acids,while the length of LvLRP1-2 is 787 bp encoding 183 amino acids.The cDNA sequences of the two transcripts are highly consistent,except for the only difference that the cDNA of LvLRP1-1 contained extra 129 bp that encodes 43 more amino acids than LvLRP1-2.Tissue expression analysis showed that LvLRP1-1 was highly expressed in the eye stalk,epidermis,brain and heart,while LvLRP1-2 was highly expressed only in the eye stalk and epidermis.The change in the transcriptional expression of LvLRP1-1 and LvLRP1-2 after Vibrio parahaemolyticus infection was consistent,and the gene expression was significantly increased at 3 hours after infection,and returned to the normal level after 6 hours.The load of Vibrio and mortality rate after Vibrio infection were significantly increased by simultaneously interfering LvLRP1-1 and LvLRP1-2 with double-stranded RNA.Therefore,LRP1 plays an important role in the resistance to Vibrio infection in L.vannamei.
关 键 词:凡纳滨对虾 低密度脂蛋白受体相关蛋白 组织表达分析 副溶血弧菌
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