在Listeria monocytogenes感染的小鼠模型中进行基因表达概况的参考基因
Lethicia Souza Tavares1, Roberta Lane Oliveira-Silva2, Marcelo Tigre Moura3
1Departamento de Biologia, Universidade Federal Rural de Pernambuco, Recife, PE, Brasil.
BioTechniques
|December 19, 2023
概括
在小鼠模型中找到正确的参考基因 (RGs) 对于准确的RT-qPCR分析至关重要. 这项研究确定了感染Listeria monocytogenes的巨细胞和脏的特定RG,改善了基因表达特征.
科学领域:
- 免疫学 免疫学 免疫学
- 分子生物学分子生物学
- 遗传学 遗传学是一种遗传学.
背景情况:
- 定量实时PCR (RT-qPCR) 对于分析基因表达是必不可少的.
- 在RT-qPCR中准确的正常化依赖于稳定的参考基因 (RG).
- 识别特定上下文的RG对于生物研究的可靠结果至关重要.
研究的目的:
- 在小鼠巨细胞 (pMØ) 和脏中识别和验证RT-qPCR的稳定参考基因.
- 评估不同RG在Listeria monocytogenes感染期间对基因表达分析的影响.
- 为了确保在感染的小鼠模型中准确的基因表达概况.
主要方法:
- 在实验室中感染小鼠腹巨细胞 (pMØ) 与L. monocytogenes.
- 在体内感染小鼠L. monocytogenes,随后收集脏.
- 从各种来源 (多种原料,文献,新设计) 选择候选的RG.
- 使用GeNorm,NormFinder,BestKeeper,Delta-CT和RefFinder算法对RG稳定性的统计分析.
主要成果:
- 对于感染的pMØ,B2m,H3f3a和Ppia被确定为最稳定的RG.
- 对于感染的脏组织,Actb,Ubc和Ppia成为最好的RGs.
- 在两种模型中,RGs的选择都影响了对细胞因子相对表达的估计.
- 在L. monocytogenes感染的小鼠模型中,上下文特定的RGs优化了基因表达概况.
结论:
- 对L. monocytogenes感染的小鼠模型需要使用RT-qPCR的特定上下文参考基因.
- 已识别的基因基因组 (pMØ的B2m-H3f3a-Ppia;的Actb-Ubc-Ppia) 提高了基因表达分析的准确性.
- 这项研究强调了严格的RG选择对于可靠的分子分析在传染病研究中的重要性.
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