|本期目录/Table of Contents|

[1]钟燕璇,郭欣悦,史小雨,等.生长素诱导的蛋白降解系统在伯氏疟原虫中的应用[J].天津医科大学学报,2025,31(05):447-452.[doi:10.20135/j.issn.1006-8147.2025.05.0447]
　ZHONG Yanxuan,GUO Xinyue,SHI Xiaoyu,et al.Application of auxin-inducible degron system in Plasmodium berghei[J].Journal of Tianjin Medical University,2025,31(05):447-452.[doi:10.20135/j.issn.1006-8147.2025.05.0447]
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生长素诱导的蛋白降解系统在伯氏疟原虫中的应用(PDF)
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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
31卷
期数:
2025年05期
页码:
447-452
栏目:
基础医学
出版日期:
2025-09-20

文章信息/Info

Title:
Application of auxin-inducible degron system in Plasmodium berghei
文章编号:
1006-8147(2025)05-0447-06
作者:
钟燕璇; 郭欣悦; 史小雨; 王倩
天津医科大学基础医学院免疫学系，天津 300070
Author(s):
ZHONG Yanxuan;  GUO Xinyue;  SHI Xiaoyu;  WANG Qian
Department of Immunology， School of Basic Medical Sciences， Tianjin Medical University， Tianjin 300070， China
关键词:
伯氏疟原虫; AID系统; YOP1
Keywords:
Plasmodium berghei;  AID system;  YOP1
分类号:
R382.3+1
DOI:
10.20135/j.issn.1006-8147.2025.05.0447
文献标志码:
A
摘要:
目的：在伯氏疟原虫中构建YOP1的二代生长素诱导蛋白降解（AID 2）系统，并降解YOP1-miniIAA7-3×Flag蛋白。方法：利用双臂同源重组构建表达AtAFB2 （F74A）-mCherry的疟原虫（Pbafb2），继而在Pbafb2疟原虫的yop1基因的C端融合miniIAA7-3×Flag，得到Pbyop1-aid疟原虫；将Pbyop1-aid疟原虫进行体外培养，分为实验组和对照组，实验组添加1 μmol/L 5-Ad-IAA，对照组添加等体积DMSO，免疫印迹检测AID 2系统对YOP1-miniIAA7-3×Flag蛋白的降解效果。结果：获得质粒AID03和AID05。获得转基因疟原虫Pbafb2和Pbyop1-aid。5-Ad-IAA处理2 h后，与对照组相比，实验组YOP1-miniIAA7-3×Flag蛋白水平显著下降（t=10.69，P<0.01）。结论：在伯氏疟原虫中成功建立了AID 2系统，可降解YOP1-miniIAA7-3×Flag蛋白。
Abstract:
Objective： To construct an auxin-inducible degron 2（AID 2） system for YOP1 in Plasmodium berghei and induce the degradation of the YOP1-miniIAA7-3×Flag fusion protein. Methods： Construction of P. berghei expressing AtAFB2 （F74A）-mCherry （Pba-fb2） was achieved using dual-arm homologous recombination. Subsequently， the miniIAA7-3×Flag were fused to the C-terminus of the yop1 gene in Pbafb2 parasites to generate Pbyop1-aid parasites. The Pbyop1-aid parasites were cultured in vitro， and divided into the experimental group and the control group. The experimental group was treated with 1 μmol/L 5-Ad-IAA， while the control group was treated with an equal volume of DMSO. Western blotting was performed to assess the degradation efficiency of YOP1-miniIAA7-3×Flag protein by the AID 2 system. Results： Plasmids AID03 and AID05 were obtained. The transgenic parasite lines Pbafb2 and Pbyop1-aid were constructed. After treatment with 5-Ad-IAA for 2 hours， the level of YOP1-miniIAA7-3×Flag protein in the experimental group was significantly reduced compared with the control group（t=10.69，P<0.01）. Conclusion： The AID 2 system is successfully established in P. berghei， and can induce the degradation of the YOP1-miniIAA7-3×Flag fusion protein.

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相似文献/References:

[1]魏超,史小雨,王倩.伯氏疟原虫红细胞膜相关蛋白1缺失突变体的构建和鉴定[J].天津医科大学学报,2022,28(02):135.
　WEI Chao,SHI Xiao-yu,WANG Qian.Construction and identification of plasmodium berghei erythrocyte membrane associated protein 1 deletion mutant[J].Journal of Tianjin Medical University,2022,28(05):135.

备注/Memo

备注/Memo:
基金项目:国家自然科学基金青年项目（32200976），国家自然科学基金面上项目（32070701）
作者简介:钟燕璇（1997-），女，硕士在读，研究方向：免疫学；通信作者：王倩，E- mail: wangq@tmu.edu.cn。
更新日期/Last Update: 2025-10-01