2023, 43(2): 18-25. doi: 10.3969/j.issn.1673-9159.2023.02.003
关键词： 凡纳滨对虾 , 过氧化物还原酶（Prx） , 基因克隆 , 脂多糖 , 4-壬基酚（4-NP）
Keywords： Litopenaeus vannamei , peroxiredoxin (Prx) , gene cloning , lipopolysaccharide , 4-nonylphenol (4-NP)
【目的】探究凡纳滨对虾（Litopenaeus vannamei）过氧化物还原酶3 （Peroxiredoxin 3，Prx3）基因（LvPrx3）的结构特征、组织分布和抗胁迫能力，为揭示凡纳滨对虾抗环境胁迫的应答机制提供理论依据。【方法】通过RACE克隆技术取得LvPrx3基因的全长cDNA序列，应用Clustal X、EditSeq、ExPASy、MEGA 6.0、SignalP 5.0和TMHMM2.0等多个软件开展Prx3的生物信息学分析。运用半定量PCR和qRT-PCR技术检测分析LvPrx3基因在多种组织和不同胁迫条件下的表达响应情况。【结果】LvPrx3基因cDNA全长序列962 bp，内含45 bp 5′端非编码区（5′-UTR）、233 bp 3′端非编码区（3′-UTR）和684 bp开放阅读框（ORF），可编码227个氨基酸残基。LvPrx3的蛋白分子质量为25.09 ku，理论等电点（pI）为6.22，属于典型的2-Cys Prx，包含高度保守的“FYPLDFTFVCPTE” N端和“GEVCPA” C端。进化树分析显示，LvPrx3与节肢动物门的Prx亲缘关系较近，与斑节对虾（Penaeus monodon）的Prx氨基酸序列相似性最高，为98%。LvPrx3在眼柄中的表达量最高，肠道次之，在其他组织中的表达量无明显区别。脂多糖（LPS）注射后，眼柄中LvPrx3的表达分别在胁迫3 h和12~48 h时显著上调，而肠道中，LvPrx3表达量在胁迫3~24 h时均显著下调；在4-壬基酚（4-NP）胁迫后，眼柄中LvPrx3表达量在胁迫6~24 h后显著上调，24 h时最高，为对照组的4.11倍；肠道中的LvPrx3表达量在胁迫12 h时显著上调，在胁迫48 h时显著下调；以上显著性水平α均为0.05。【结论】LvPrx3基因属典型的2-Cys Prx，主要在凡纳滨对虾眼柄和肠道中高表达。脂多糖LPS刺激和4-NP胁迫可明显诱导凡纳滨对虾LvPrx3基因的表达水平，表明LvPrx3在对虾抵御病原菌感染及抗逆境胁迫的调控中发挥重要作用。
【目的】探究凡纳滨对虾（Litopenaeus vannamei）过氧化物还原酶3 （Peroxiredoxin 3，Prx3）基因（LvPrx3）的结构特征、组织分布和抗胁迫能力，为揭示凡纳滨对虾抗环境胁迫的应答机制提供理论依据。【方法】通过RACE克隆技术取得LvPrx3基因的全长cDNA序列，应用Clustal X、EditSeq、ExPASy、MEGA 6.0、SignalP 5.0和TMHMM2.0等多个软件开展Prx3的生物信息学分析。运用半定量PCR和qRT-PCR技术检测分析LvPrx3基因在多种组织和不同胁迫条件下的表达响应情况。【结果】LvPrx3基因cDNA全长序列962 bp，内含45 bp 5′端非编码区（5′-UTR）、233 bp 3′端非编码区（3′-UTR）和684 bp开放阅读框（ORF），可编码227个氨基酸残基。LvPrx3的蛋白分子质量为25.09 ku，理论等电点（pI）为6.22，属于典型的2-Cys Prx，包含高度保守的“FYPLDFTFVCPTE” N端和“GEVCPA” C端。进化树分析显示，LvPrx3与节肢动物门的Prx亲缘关系较近，与斑节对虾（Penaeus monodon）的Prx氨基酸序列相似性最高，为98%。LvPrx3在眼柄中的表达量最高，肠道次之，在其他组织中的表达量无明显区别。脂多糖（LPS）注射后，眼柄中LvPrx3的表达分别在胁迫3 h和12~48 h时显著上调，而肠道中，LvPrx3表达量在胁迫3~24 h时均显著下调；在4-壬基酚（4-NP）胁迫后，眼柄中LvPrx3表达量在胁迫6~24 h后显著上调，24 h时最高，为对照组的4.11倍；肠道中的LvPrx3表达量在胁迫12 h时显著上调，在胁迫48 h时显著下调；以上显著性水平α均为0.05。【结论】LvPrx3基因属典型的2-Cys Prx，主要在凡纳滨对虾眼柄和肠道中高表达。脂多糖LPS刺激和4-NP胁迫可明显诱导凡纳滨对虾LvPrx3基因的表达水平，表明LvPrx3在对虾抵御病原菌感染及抗逆境胁迫的调控中发挥重要作用。
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