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首页> 外文期刊>Journal of Veterinary Research >Construction and activity analyses of single functional mouse peroxiredoxin 6 (Prdx6)
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Construction and activity analyses of single functional mouse peroxiredoxin 6 (Prdx6)

机译：单函数小鼠过氧杂志6（PRDX6）的构建和活性分析

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摘要

Introduction: Peroxiredoxin 6 (Prdx6) is a bifunctional protein with glutathione peroxidase activity and phospholipase A2 activity. Previous studies have shown a significant positive correlation between the intracellular survival ability of Brucella and Prdx6. Here, the Prdx6 enzyme with a single activity was constructed to facilitate study of the relationship between the single function of Prdx6 and Brucella infection. Material and Methods: The target open reading frame (ORF) DNAs of Prdx6 with a single active centre were prepared using gene splicing by overlap extension PCR (SOE-PCR), and the recombinant eukaryotic expression plasmids inserted by Prdx6 with the single activity centre were constructed and transfected into murine Raw264.7 macrophages. The glutathione peroxidase activity and phospholipase A2 activity of the constructed Prdx6 were examined. Results: The core centres (Ser(32) and Cys(47)) of Prdx6 were successfully mutated by changing the 94th nucleotide from T to G and the 140th nucleotide from G to C in the two enzyme activity cores, respectively. The constructed recombinant plasmids of Prdx6 with the single active centre were transfected into murine macrophages showing the expected single functional enzyme activity, which MJ33 or mercaptosuccinate inhibitors were able to inhibit. Conclusion: The constructed mutants of Prdx6 with the single activity cores will be a benefit to further study of the biological function of Prdx6 with different enzyme activity.
机译：简介：过氧化毒素6（PRDX6）是一种双官能蛋白，具有谷胱甘肽过氧化物酶活性和磷脂酶A2活性。先前的研究表明，布鲁氏菌和PRDX6的细胞内存活能力之间存在显着的正相关性。这里，构建了具有单一活性的PRDX6酶以促进PRDX6和Brucella感染的单一功能之间的关系。材料和方法：使用通过重叠延伸PCR（SOE-PCR）的基因剪接制备具有单个活性中心的PRDX6的靶开放阅读框（ORF）DNA，并通过PRDX6与单一活动中心插入的重组真核表达质粒是构建并转染到鼠Raw264.7巨噬细胞中。检查了构建的PRDX6的谷胱甘肽过氧化物酶活性和磷脂酶A2活性。结果：PRDX6的核心中心（SER（32）和CYS（47））通过分别将来自T至G的第94个核苷酸与G至C中的第140个核苷酸从G至C中改变为两种酶活性核来成功突变。将PRDX6的构建重组质粒与单一活性中心转染到鼠巨噬细胞，显示预期的单一官能酶活性，MJ33或巯基琥珀酸抑制剂能够抑制。结论：具有单次活性核心的PRDX6的构建突变体将是进一步研究PRDX6具有不同酶活性的生物学功能的有益。

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  • 来源
    《Journal of Veterinary Research》 |2019年第1期|共7页
  • 作者

    Wang Lu-Lu; Lu Shi-Ying; Hu Pan; Fu Bao-Quan; Li Yan-Song; Zhai Fei-Fei; Ju Dan-Di; Zhang Shi-Jun; Su Bing; Zhou Yu; Liu Zeng-Shan; Ren Hong-Lin;

  • 作者单位

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Chinese Acad Agr Sci State Key Lab Vet Etiol Biol Key Lab Vet Publ Hlth Minist Agr Key Lab Vet Parasitol Gansu Prov Lanzhou Vet Res Lanzhou 730046 Gansu Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

    Jilin Univ Inst Zoonosis Key Lab Zoonosis Res Coll Vet Med Xi An Du Lu 5333 Changchun 130062 Jilin Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 动物医学（兽医学）;
  • 关键词

    mouse; peroxiredoxin 6; glutathione peroxidase; phospholipase A2; SOE-PCR;

    机译：小鼠;过氧杂志6;谷胱甘肽过氧化物酶;磷脂酶A2;SOE-PCR;

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