嗜水气单胞菌刺激下鲤血清质子转运载体15a2 (Slc15a2)的表达分析
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Q 78; S 941.4

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国家自然科学基金(31801032); 淡水鱼类育种国家地方联合工程实验室开放课题(KF-2016-03);河南师范大学博士基金(qd16159)


Expression analysis of serum Slc15a2 in common carp (Cyprinus carpio) stimulated by Aeromonas hydrophila
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National Natural Science Foundation of China (31801032); Open project of National and Local Joint Engineering Laboratory for Freshwater Fish Breeding (KF-2016-03); Ph.D. Foundation of Henan Normal University (qd16159)

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    摘要:

    为解析鲤质子转运载体15a2 (Slc15a2)在嗜水气单胞菌侵染过程中对机体免疫应答机制的影响,实验通过制备鼠源Slc15a2-1和Slc15a2-2多克隆抗体,采用酶联免疫吸附测定 (ELISA) 技术检测嗜水气单胞菌感染后鲤血清中Slc15a2-1和Slc15a2-2蛋白表达变化。结果显示,①感染后2个拷贝的最高蛋白表达水平与空白对照组相比,分别增加了4.81和2.48倍;与Slc15a2-1相比Slc15a2-2的表达量从0~48 h一直处于较高表达水平。②从整体来看,Slc15a2的2个拷贝具有相似的表达趋势,均为先升高后降低、再升高再降低;但Slc15a2-1在3 h达到第1个表达高峰,而Slc15a2-2则在6 h达到第1个表达高峰。③ 6~24 h的4个时期中,Slc15a2-1和Slc15a2-2蛋白表达呈现互补的表达趋势,即在同一时期一个拷贝的表达量显著下降,另一个拷贝的表达量显著升高,但总体维持较高表达水平。结果表明,本实验制备的鼠源Slc15a2-1和Slc15a2-2多克隆抗体具有较高的亲和力和特异性,当鲤受到嗜水气单胞菌侵染后,2个拷贝整体具有相似的表达趋势,说明可能具有相似的基因功能;与Slc15a2-1相比,Slc15a2-2在0~48 h之间具有显著的高表达量,推测该基因为2个拷贝中的主效基因,并且6~24 h时,2个拷贝一直呈现互补的表达方式,导致Slc15a2表达量维持在高水平,面对病原侵害时能做有效响应。本研究为深入了解鲤Slc15a2的免疫防御机制提供一定的研究基础。

    Abstract:

    To analyze the effect of the Cyprinus carpio Slc15a2 gene on the immune response mechanism during Aeromonas hydrophila infestation, this experiment prepared rat-derived Slc15a2-1 and Slc15a2-2 polyclonal antibodies, and changes in protein expression of Slc15a2-1 and Slc15a2-2 in C. carpio sera following A. hydrophila infection was detected by enzyme linked immunosorbent assay (ELISA). The results showed that ① compared with the blank control the maximum protein expression levels of the two copies after infection were increased by 4.81 and 2.48 times, respectively. The protein expression of Slc15a2-2 was consistently higher from 0 h to 48 h compared to that of Slc15a2-1. ② Overall, there was a similar expression trend between the two copies of Slc15a2, with both increasing, then decreasing, then increasing and then decreasing again. However, Slc15a2-1 reached its first expression peak at 3 h, while Slc15a2-2 reached its first expression peak at 6 h. ③ Slc15a2-1 and Slc15a2-2 protein expression showed complementary expression trends during the four periods from 6 h to 24 h. That is, the expression of one copy decreased significantly and the expression of the other copy increased significantly during the same period, but the overall expression level remained high. The results indicate that the two polyclonal antibodies prepared in this experiment have high affinity and specificity, and the overall similar expression trend between the two copies after infection with A. hydrophila suggests that they may have similar gene functions. Slc15a2-2 had significantly higher expression between 0 h and 48 h than Slc15a2-1, possibly as the major gene in the copy. The two copies showed complementary expression from 6 h to 24 h, and the co-expression was maintained at a high level, enabling an effective response in the face of pathogen attack. This study may provide some basis for insight into the regulatory immune response mechanism of C. carpio Slc15a2 gene.

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姜洲,薛亚果,张猛,朱雷,王磊,李学军,董传举.嗜水气单胞菌刺激下鲤血清质子转运载体15a2 (Slc15a2)的表达分析[J].水产学报,2022,46(5):797~804

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历史
  • 收稿日期:2020-11-26
  • 最后修改日期:2021-06-08
  • 录用日期:2021-08-14
  • 在线发布日期: 2022-05-31
  • 出版日期: 2022-05-01