鳜KLF6序列特征、组织表达及其对BCAAs代谢酶基因表达的影响  

Sequence Characteristics and Tissue Expression and Its Effect on Gene Expression of BCAAs Metabolic Enzymes of KLF6 Gene in Siniperca chuatsi

作　　者：宾琴 宾石玉[1,2] 包凌晟 褚武英[3,4] 张建社[3,4] 程聪益[3,4] 曾维 姚舟 BIN Qin;BIN Shiyu;BAO Lingsheng;CHU Wuying;ZHANG Jianshe;CHENG Congyi;ZENG Wei;YAO Zhou(College of Life Sciences,Guangxi Normal University,Guilin Guangxi 541006,China;Key Laboratory of Ecology and Environmental Protection of Rare and Endangered Plants and Animals(Guangxi Normal University),Ministry of Education,Guilin Guangxi 541006,China;School of Biological and Chemical Engineering,Changsha University,Changsha Hunan 410022,China;Hunan Provincial Key Laboratory of Aquatic Animal Nutrition and Quality Control(Changsha University),Changsha Hunan 410022,China)

机构地区：[1]广西师范大学生命科学学院,广西桂林541006 [2]珍稀濒危动植物生态与环境保护教育部重点实验室(广西师范大学),广西桂林541006 [3]长沙学院生物与化学工程学院,湖南长沙410022 [4]水生动物营养与品质调控湖南省重点实验室(长沙学院),湖南长沙410022

出　　处：《广西师范大学学报（自然科学版）》2024年第6期236-243,共8页Journal of Guangxi Normal University:Natural Science Edition

基　　金：国家自然科学基金(31560718,31820103016)。

摘　　要：为研究鳜KLF6基因序列特征、组织表达及其对支链氨基酸代谢酶基因表达的影响,通过在线工具和软件对KLF6基因进行生物信息学分析,运用实时荧光定量PCR(RT-qPCR)技术,检测KLF6基因在鳜7个不同组织的表达丰度及用siRNA干扰后KLF6基因和BCAAs代谢相关酶基因的mRNA表达量。结果表明:鳜KLF6基因编码区全长为3791 bp(登录号:XM_044176560.1),其中5′端和3′端分别为271和2659 bp,开放阅读框860 bp,共编码氨基酸286个。与鳃棘鲈KLF6基因的亲缘关系最近(98.6%),与鲤鱼较远(82.3%)。KLF6有1个KLF6-N型结构域(2-204 aa)、1个COG5048结构域(194-268 aa)和3个Znf-C2H2结构域(分别位于203-227、233-257、263-285个氨基酸之间)。鳜KLF6蛋白二级结构由无规则卷曲(56.99%)、α-螺旋(26.22%)、延伸链(10.84%)和β-转角(5.94%)相互连接而成。RT-qPCR结果发现,KLF6基因在鳜组织的相对表达量大小由高到低依次为心脏、脾脏、脑、肾脏、肠道、肌肉、肝脏。其中,心脏的表达量显著高于肝脏(P<0.01)和肌肉(P<0.05),脾脏、脑、肾脏及肠道的表达量高于肝脏(P<0.05)。在KLF6基因的mRNA表达量受到抑制后,BCAAs代谢酶(BCKDHb、ACADM、MCCC1、MCCC2、MUT、IVD)基因的mRNA表达量显著上升(P<0.05)。本研究为KLF6基因在鳜肌肉BCAAs代谢的调控机制研究提供了依据。In order to study the sequence characteristics and tissue expression of KLF6 gene in Siniperca chuatsi and its influence on the expression of branchable amino acid metabolizing enzyme gene,bioinformatics analysis of KLF6 gene was conducted through online tools and software.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression abundance of KLF6 gene in 7 different tissues of the Sinfinfish and the mRNA expression levels of KLF6 gene and BCAAs metabolizing enzyme related genes after siRNA interference.The results showed that the total length of the coding region of the KLF6 gene of S.chuatsi was 3791 bp(entry number:XM_044176561.1),in which the 5′end and 3′end were 271 bp and 2659 bp,respectively,and the open reading frame was 860 bp,encoding a total of 286 amino acids.It was most closely related to the KLF6 gene(98.6%)and more distant to the KLF6 gene(82.3%)of carp.KLF6 has one KLF6-N type domain(2-204 aa),one COG5048 domain(194-268 aa)and three Znf-C2H2 domains,which were located between 203-227,233-257 and 263-285 amino acids,respectively.The structure of the S.chuatsi KLF6 protein was composed of random curl(56.99%),α-helix(26.22%),extension chain(10.84%)andβ-angle(5.94%).The results of RT-qPCR showed that the relative expression level of KLF6 gene in S.chuatsi tissues was in the following order from high to low:heart,spleen,brain,kidney,intestine,muscle and liver,and the expression level in the center was significantly higher than that in liver(P<0.01)and muscle(P<0.05),while the expression level in spleen,brain,kidney and intestine was higher than that in liver(P<0.05).The mRNA expression of BCAAs metabolizing enzymes(BCKDHb,ACADM,MCCC1,MCCC2,MUT,IVD)was significantly increased after the mRNA expression of KLF6 gene was inhibited(P<0.05).This study provides a basis for the study of the regulatory mechanism of KLF6 gene in muscle BCAAs metabolism of S.chuatsi.

关 键 词：鳜 转录因子 支链氨基酸 代谢酶 组织表达 

分 类 号：S917.4[农业科学—水产科学]