CAJ | 학술논문

ATG5(Autophagy related gene 5)基因是一种自噬相关基因,是形成自噬体的重要基因, ATG5基因的表达对自噬调控的成熟有重要作用。本研究中通过克隆,首次得到太平洋鳕Gadus macrocephalus ATG5 cDNA部分序列,该序列长为895 bp,含有完整的开放阅读框( Open reading frame, ORF),编码275个氨基酸,其编码的蛋白质相对分子量约为32200；经核苷酸序列比对发现,太平洋鳕ATG5核苷酸序列与半滑舌鳎Cynoglossus semilaevis、尼罗罗非鱼Oreochromis niloticus ATG5核苷酸序列的相似性均高达87.27%；利用太平洋鳕与其他物种ATG5核苷酸序列构建系统进化树,结果显示, ATG5核苷酸序列系统进化树反映的亲缘关系基本符合传统分类学观点；氨基酸序列分析结果显示, ATG5蛋白氨基酸序列具有较高的保守性；利用绝对荧光定量PCR方法对ATG5基因在太平洋鳕各组织中的转录水平进行检测,结果显示,肝、脾和肾中的表达量高于其他组织,鳃、肾、脾、脑、肝和肌肉各组织中的 ATG5 mRNA 拷贝数分别为11.61708、20.44925、19.70687、4.83984、27.43497、2.95475 copies/ng。本研究结果将为进一步研究太平洋鳕细胞吞噬机制奠定基础,也为太平洋鳕的免疫机理和相关研究提供基础数据。
ATG5(Autophagy related gene 5)기인시일충자서상관기인,시형성자서체적중요기인, ATG5기인적표체대자서조공적성숙유중요작용。본연구중통과극륭,수차득도태평양설Gadus macrocephalus ATG5 cDNA부분서렬,해서렬장위895 bp,함유완정적개방열독광( Open reading frame, ORF),편마275개안기산,기편마적단백질상대분자량약위32200；경핵감산서렬비대발현,태평양설ATG5핵감산서렬여반활설탑Cynoglossus semilaevis、니라라비어Oreochromis niloticus ATG5핵감산서렬적상사성균고체87.27%；이용태평양설여기타물충ATG5핵감산서렬구건계통진화수,결과현시, ATG5핵감산서렬계통진화수반영적친연관계기본부합전통분류학관점；안기산서렬분석결과현시, ATG5단백안기산서렬구유교고적보수성；이용절대형광정량PCR방법대ATG5기인재태평양설각조직중적전록수평진행검측,결과현시,간、비화신중적표체량고우기타조직,새、신、비、뇌、간화기육각조직중적 ATG5 mRNA 고패수분별위11.61708、20.44925、19.70687、4.83984、27.43497、2.95475 copies/ng。본연구결과장위진일보연구태평양설세포탄서궤제전정기출,야위태평양설적면역궤리화상관연구제공기출수거。
ATG5 as an abbreviation of autophagy related gene 5 is an important gene for the formation of autophago-some, and plays a vital role in the maturation of autophagy regulation. In this study, a cDNA sequence of ATG5 was first cloned in Pacific cod Gadus macrocephalus using gene cloning, and was found to be 895 bp encoding 275 amino acid residues with molecular weight of about 32 200 in length with a whole open reading frame( ORF) . The nucleotide sequence alignment showed that there was as high as similarity of 87. 27% in ATG5 among Pacific cod, Nile tilapia Oreochromis niloticus, and half-smooth tongue-sole Cynoglossus semilaev. The phylogenetic tree con-structed by ATG5 nucleotide sequences of Pacific cod and some other animals reflected a genetic relationship which conformed to the viewpoint of traditional taxonomy. The amino acid sequence analysis revealed that ATG5 had high conservation. The expression of ATG5 in various tissues of Pacific cod using relative quantitative PCR and the abso-lute quantitative PCR showed that the higher expression levels of ATG5 mRNA were observed in liver, spleen and kidney than in other tissues, with 11. 617 08 copies/ng in gill, 20. 449 25 copies/ng in kidney, 19. 706 87 copies/ng in spleen, 4. 839 84 copies/ng in brain, 27. 434 97 copies/ng in liver, and 2. 954 75 copies/ng in muscle. The findings lay the foundation for further study of the phagocytosis mechanism and provide basic data with understand-ing of immune mechanisms in Pacific cod.