中华倒刺鲃β-淀粉样前体蛋白的克隆及表达分析  

Cloning and Expression Analysis of β -amyloidPrecursor Protein from SpinibarbusSinensis

作　　者：雷昊明 赵月桐 周婵娟 蔡志宏 李俊 Lei Haoming;Zhao Yuetong;Zhou Chanjuan;Cai Zhihong;Li Jun(School of Biological and Food Engineering,Huaihua University,Huaihua 418000,China;Hunan Key Laboratory of Source Research and Utilization of Ethnic Medicinal Plants,Huaihua 418000,China)

机构地区：[1]怀化学院生物与食品工程学院,湖南怀化418000 [2]民族药用植物资源研究与利用湖南省重点实验室,湖南怀化418000

出　　处：《黑龙江科学》2024年第24期98-101,105,共5页Heilongjiang Science

基　　金：湖南省大学生创新性训练项目(S202010548069);湖南省教育厅青年项目(21B0700)。

摘　　要：β-淀粉样前体蛋白(amyloid beta precursor protein, APP)作为淀粉样β肽形成的关键蛋白,在阿尔茨海默病的形成过程中发挥了重要作用,但其在鱼类免疫系统中的功能还未被探究。利用聚合酶链式反应技术克隆出中华倒刺鲃(SpinibarbusSinensis)APP基因的cDNA编码序列(GenBank编号PP716090),生物信息学分析表明该序列全长2220 bp,共编码739个氨基酸,APP蛋白存在A4_EXTRA、KU、Coiled coil和Transmembrane结构域,邻接法构建的系统进化树显示中华倒刺鲃的APP与露斯塔野鲮的APP聚类在一起。将APP构建到pcDNA3.1-MCS-3xflag载体中进行双酶切检测及测序检测,结果表明表达载体构建成功。利用RT-qPCR技术分析APP的表达模式,结果表明APP基因在中华倒刺鲃的脑、心脏、肌肉、肾脏、眼睛、尾鳍和脾脏中均有表达,且在脾脏中的相对表达量最高。此研究为进一步探讨中华倒刺鲃APP基因功能提供了基础。As a key protein in the formation of amyloidβprecursor protein,amyloid beta precursor protein(APP)plays an important role in the formation of Alzheimer’s disease,but its function in the immune system of fish has not been explored.The study clones cDNA coding sequence(GenBank number PP716090)of the APP gene of SpinibarbusSinens by polymerase chain reaction(PCR).Bioinformatics analysis shows that the sequence is 2220 bp in length and encoded 739 amino acids.APP protein exists in A4_EXTRA,KU,Coiled coil and Transmembrane.The phylogenetic tree constructed by adjacency method show that APP of SpinibarbusSinensis and APP of Labeo Lustaensis are clustered together.The study conducts APP into pcDNA3.1-MCS-3xflag vector for double enzyme digestion detection and sequencing detection.The results show that the expression vector is successfully constructed.The study analyzes the expression pattern of APP by RT-qPCR.The results show that APP gene is expressed in brain,heart,muscle,kidney,eye,tail fin and spleen.The relative expression level is the highest in spleen.This study provides a basis for further study on the function of the APP gene in SpinibarbusSinensis.

关 键 词：Β-淀粉样前体蛋白 鱼类免疫系统 中华倒刺鲃 分子生物学技术 

分 类 号：Q958.8[生物学—动物学]