克氏原螯虾<i>traf</i>6<i> like</i>基因参与抗细菌先天免疫反应的研究

蔺思函; 杨兵兵; 李倩倩; 沈秀丽; 杜志强

doi:10.19303/j.issn.1008-0384.2022.010.001

克氏原螯虾traf6 like基因参与抗细菌先天免疫反应的研究

doi: 10.19303/j.issn.1008-0384.2022.010.001

1.
内蒙古科技大学生命科学与技术学院，内蒙古　包头　014010
2.
内蒙古科技大学图书馆，内蒙古　包头 014010

基金项目: 内蒙古自治区自然科学基金项目（2020JQ03）；国家自然科学基金项目（32060834）；内蒙古自治区研究生科研创新项目（S20210160Z）

详细信息

作者简介:
蔺思函（1998−），女，硕士研究生，研究方向：无脊椎动物的先天免疫（E-mail：2836019366@qq.com）

通讯作者:
杜志强（1980−），男，博士，教授，研究方向：无脊椎动物的先天免疫（E-mail：nmdzq@163.com）

中图分类号: Q 786
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出版历程
- 收稿日期: 2022-01-21
- 修回日期: 2022-04-02
- 刊出日期: 2022-10-30

Molecular Mechanism of traf6 like Gene in Innate Immunity of Procambarus clarkii

1.
College of Life Science and Technology, Inner Mongolia University of Science and Technology, Baotou, Inner Mongolia　014010, China
2.
Library, Inner Mongolia University of Science and Technology, Baotou, Inner Mongolia　014010, China

摘要

摘要: 目的深入研究克氏原螯虾（Procambarus clarkia）traf6 like基因参与先天免疫调节的功能。方法以Pc-traf6 like基因为研究对象，利用RNAi技术干扰目的基因表达量之后，进行细菌模拟刺激，并且通过实时荧光定量PCR（qRT-PCR）检测细胞凋亡相关基因表达量的变化情况。结果干扰Pc-traf6 like基因，通过干扰效果检测，显示48 h时干扰效果良好，可用于后续试验。在目的基因干扰后，进行金黄色葡萄球菌（Staphylococcus aureus）刺激，检测表明促凋亡基因Bax、抗凋亡基因Bcl 2、细胞凋亡关键基因Caspase 3不同程度上调表达；在目的基因干扰后，进行鲶爱德华氏菌（Edwardsiella ictaluri ）刺激，检测表明促凋亡基因Bax、抗凋亡基因Bcl 2、细胞凋亡关键基因Caspase 3不同程度上调表达。结论 Pc-traf6 like基因被干扰，机体在受到金黄色葡萄球菌及鲶爱德华氏菌刺激后，Bax、Bcl 2和Caspase 3基因的相对表达量上调，推测Pc-traf6 like基因通过调控细胞凋亡参与了先天免疫反应。
- 克氏原螯虾 /
- RNA干扰 /
- 细胞凋亡 /
- 先天免疫
Abstract: Objective Role of traf6 like gene in innate immunity regulation of Procambarus clarkia(Pc-traf6 like) was investigated. Methods RNAi technology was employed to interfere the expression of Pc-traf6 like gene. Bacterial simulation was performed to determine the changes of apoptosis-related gene expression by real-time quantitative PCR (qRT-PCR). Result The interference on the gene was effective after 48 h for the subsequent experiments. The interference-induced pro-apoptotic gene Bax, anti-apoptotic gene Bcl 2, and apoptotic protease Caspase 3 were upregulated in varying degrees with the stimulation of Staphylococcus aureus or Edwardsiella ictaluri. Conclusion After the Pc-traf6 like gene was effectively interfered, the relative expressions of Bax, Bcl 2, and Caspase 3 were upregulated after the P. clarkia was stimulated by either S. aureus or E. ictaluri indicating an involvement of Pc-traf6 like gene in the innate immune response by the regulating apoptosis.
- Procambarus clarkii /
- RNA interference /
- apoptosis /
- innate immunity

HTML全文

图 1 Pc-traf6 like基因双链RNA电泳

M：标准DNA Marker；1～2：目的基因双链RNA条带。

Figure 1. Electrophoresis on double-stranded RNA validation of Pc-traf6 like gene

M: standard DNA marker; 1–2: target gene double-stranded RNA bands.

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图 2 Pc-traf6 like基因双链RNA干扰效果检测

18S RNA作为内参对照。不同字母表示不同时间点同一处理组差异显著（P＜0.05）；*、**表示同一时间点不同处理组之间差异显著（P＜0.05）或极显著（P＜0.01）。图3、4同。

Figure 2. Detection of interference on double-stranded RNA of Pc-traf6 like gene

18S RNA was used as inner control; data with different letters indicate significant difference in same group at times (P＜0.05); * or ** indicates significant difference between treatment groups at 0.05 or 0.01 level. Same for Figs. 3 and 4.

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图 3 金黄色葡萄球菌刺激后细胞凋亡相关基因的表达分析

A为Bax基因；B为Bcl 2基因；C为Caspase 3基因。图4同。

Figure 3. Expressions of apoptosis-related genes after S. aureus stimulation

A: Bax gene; B: Bcl gene; C: Caspase 3 gene. Same for Fig.4.

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图 4 鲶爱德华氏菌刺激后细胞凋亡相关基因的表达模式

Figure 4. Expressions of apoptosis-related genes after E. ictaluri stimulation

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