CAJ | 학술논문

微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3，LC3）是哺乳动物细胞中酵母自噬相关基因8（ATG8）的同源物，其 LC3B亚型的表达强度与自噬泡数量的多少成正相关，为自噬发生的标志蛋白之一。应用 RACE-PCR技术克隆了婺源荷包红鲤 LC3B基因的 cDNA全长序列，用 RT-PCR方法检测了该基因在荷包红鲤主要组织中的表达，并用Real-time quantitative PCR方法检测了该基因在镉胁迫下转录水平的变化，以初步揭示该基因在镉胁迫下的作用。结果表明，荷包红鲤 LC3B基因 cDNA全长为2138 bp，其中开放阅读框长378 bp，编码125个氨基酸，预测分子量为14．73 kDa，等电点为8．76。序列比对显示，其编码的氨基酸序列和斑马鱼LC3B具有最高的一致性和相似性，并在系统发育树上聚为一支。该基因在荷包红鲤肌肉、脑、鳃、脾脏、肝脏、血液、头肾、肾脏和心脏组织中均有表达；在镉胁迫下，镉诱导显著影响了该基因在肾脏组织中的转录水平。LC3 B基因参与的自噬反应可能参与了抗镉毒性作用。
미관상관단백1경련3（microtubule-associated protein 1 light chain 3，LC3）시포유동물세포중효모자서상관기인8（ATG8）적동원물，기 LC3B아형적표체강도여자서포수량적다소성정상관，위자서발생적표지단백지일。응용 RACE-PCR기술극륭료무원하포홍리 LC3B기인적 cDNA전장서렬，용 RT-PCR방법검측료해기인재하포홍리주요조직중적표체，병용Real-time quantitative PCR방법검측료해기인재력협박하전록수평적변화，이초보게시해기인재력협박하적작용。결과표명，하포홍리 LC3B기인 cDNA전장위2138 bp，기중개방열독광장378 bp，편마125개안기산，예측분자량위14．73 kDa，등전점위8．76。서렬비대현시，기편마적안기산서렬화반마어LC3B구유최고적일치성화상사성，병재계통발육수상취위일지。해기인재하포홍리기육、뇌、새、비장、간장、혈액、두신、신장화심장조직중균유표체；재력협박하，력유도현저영향료해기인재신장조직중적전록수평。LC3 B기인삼여적자서반응가능삼여료항력독성작용。
LC3(microtubule-associated protein 1 light chain 3),located in autophagosome membranes after processing,serves as a major autophagy marker.In this study,we isolated its LC3B isform from common carp (cyprinus carpio),one of the most important industrial aquatic animals in China using rapid amplifica-tion of cDNA ends (RACE).The cDNA of LC3B gene is 2138-nt in length,containing a 378-nt open read-ing frame.The putative LC3B encodes a peptide of 125 amino acids,with a calculated molecular weight of 14.73 kDa and an isoelectric point of 8.76.Multiple sequence alignment showed that the deduced amino acid sequence of common carp LC3B had the highest sequence identity (95.2%)and similarity (96.8%)to that of zebrafish.In a phylogenetic tree based on amino acid sequences from fish,mammalian,avian and am-phibian LC3 genes,the LC3 proteins from different species were divided into three subgroups (LC3A, LC3B and LC3C).The common carp LC3B formed a phylogenetic cluster with zebrafish,then other verte-brates.The mRNA transcript level of LC3B was detected in muscle,brain,gills,spleen,liver,blood,head kidney,kidney and heart tissues by RT-PCR analysis.LC3B mRNA transcript level in kidney was found significantly elevated after exposure to cadmium.This was markedly affected by exposure concentration, not by exposure time.