南方农业学报2023，Vol.54Issue(12)：3745-3753,9.DOI:10.3969/j.issn.2095-1191.2023.12.029
陆川猪ACOX1基因不同亚型克隆及其表达差异分析
Gening cloning and bioinformatics analysis of different subtypes of ACOX1 gene in Luchuan pigs
摘要
Abstract
[Objective]This study was aimed to clone different isoforms of Acyl-Coa oxidase 1(Acyl-CoA oxidase 1,ACOX1)gene in Luchuan pig and conduct bioinformatics analysis,analyze the expression of ACOX1 gene and its iso-forms in longissimus dorsi muscle of different breeds of pigs.So as to provide the basics for further exploring the regula-tory roles of ACOX1 gene in Luchuan pigs.[Method]Based on pig ACOX1 gene sequence published in GenBank(Acces-sion No(NM_001101028:1)and the CDS sequence of the Luchuan pig ACOX1 gene,The specific primers and validation primers were designed to amplify ACOX1 gene from Luchuan pig using RT-PCR technique.Total RNA was extracted from the liver of Luchuan pig aged 0 months and the longissimus dorsi muscle of Luchuan pig,Duroc,Dulu and Ludu pigs aged 8 months by TRIzol method.The relative expression of ACOX1 gene and its subtypes in longissimus dorsi muscle of Lu-chuan pig,Duroc,Dulu pig and Luchuan pig were detected by Real-time quantitative PCR.[Result]There are two subtype structures of ACOX1 gene in Luchuan pigs,the GenBank accession numbers were OQ701687 and OQ701688.the full Length sequence of ACOX1 gene CDS in in Luchuan pigs were 1986 bp,encoding 661 amino acids,the two isomers mainly undergo selective cleavage between sequences between positions 270 to 429,The molecular weight of Luchuan pig ACOX1-I was 74786.5 Da,and its theoretical isoelectric point(pI)was 7.36;The molecular weight of ACOX1-II was 74670.49 Da,and its theoretical isoelectric point(pI)was 8.17.The secondary structure of ACOX1-I protein In Luchuan pig was composed of Alpha helix(47.96%),Beta turn(10.14%),extended Chain(14.52%)and random coil(27.38%).The secondary structure of ACOX1-II protein in Luchuan pig was composed of Alpha helix(47.66%),Beta turn(10.89%),ex-tended Chain(14.37%)and random coil(27.08%).The ACOX1 protein of Luchuan pig had no transmembrane domain.The expression of ACOX1-I gene in the longissimus dorsi muscle of Luchuan pigs was the highest,but the difference was not significant(P>0.05).The expression of ACOX1-II gene in the longissimus dorsi muscle of Luchuan pig and Duroc pigs were significantly higher than other breeds(P<0.05).[Conclusion]Two different subtypes of ACOX1 gene in Lu-chuan pig were successfully cloned,,the expression of ACOX1 gene and its subtypes in the longissimus dorsi muscle of different pig breeds were significant differences.关键词
陆川猪/ACOX1基因/亚型/生物信息学分析/表达分析Key words
Luchuan pig/ACOX1gene/isoforms/bioinformatics analysis/expression analysis分类
农业科技引用本文复制引用
陈钊,陈宝剑,潘鹏丞,潘星辰,陈少梅,覃兆鲜,吴永绍,覃倩,农素群,谢炳坤..陆川猪ACOX1基因不同亚型克隆及其表达差异分析[J].南方农业学报,2023,54(12):3745-3753,9.基金项目
广西重点研发计划项目(桂科AB21196060) （桂科AB21196060）
中央引导地方科技发展专项(桂科ZY21195052) （桂科ZY21195052）
广西农业科技自筹经费项目(Z2022119,Z2022111) Guangxi Key Technologies R&D Program(Guike AB21196060) （Z2022119,Z2022111）
Central Guiding Local Science and Technology Development Special Project(Guike ZY21195052) （Guike ZY21195052）
Guangxi Agricultural Science and Technology Self fi-nancing Project(Z2022119,Z2022111). （Z2022119,Z2022111）
