The Identification of the Banana Endogenous Reference Gene MaSPS1 and the Construction of Qualitative and Quantitative PCR Detection Methods
文献类型： 外文期刊
作者： Zhu, Lili 1 ; Lin, Ying 1 ; Yang, Wenli 1 ; Pan, Zhiwen 1 ; Chen, Weiting 1 ; Yao, Juan 1 ; Sheng, Ou 3 ; Zhou, Lingyan 2 ; Jiang, Dagang 1 ;
作者机构： 1.South China Agr Univ, Coll Life Sci, Guangzhou 510642, Peoples R China
2.Zhongkai Univ Agr & Engn, Coll Agr & Biol, Guangzhou 510225, Peoples R China
3.Guangdong Acad Agr Sci, Inst Fruit Tree Res, Guangzhou 510640, Peoples R China
关键词： banana (Musa paradisiaca); endogenous reference gene; MaSPS1; qPCR
期刊名称：GENES （ 影响因子：3.5； 五年影响因子：3.9 ）
ISSN：
年卷期： 2023 年 14 卷 12 期
页码：
收录情况： SCI
摘要： Endogenous reference genes play a crucial role in the qualitative and quantitative PCR detection of genetically modified crops. Currently, there are no systematic studies on the banana endogenous reference gene. In this study, the MaSPS1 gene was identified as a candidate gene through bioinformatics analysis. The conservation of this gene in different genotypes of banana was tested using PCR, and its specificity in various crops and fruits was also examined. Southern blot analysis showed that there is only one copy of MaSPS1 in banana. The limit of detection (LOD) test showed that the LOD of the conventional PCR method is approximately 20 copies. The real-time fluorescence quantitative PCR (qPCR) method also exhibited high specificity, with a LOD of approximately 10 copies. The standard curve of the qPCR method met the quantitative requirements, with a limit of quantification (LOQ) of 1.14 x 10(-2) ng-about 20 copies. Also, the qPCR method demonstrated good repeatability and stability. Hence, the above results indicate that the detection method established in this study has strong specificity, a low detection limit, and good stability. It provides a reliable qualitative and quantitative detection system for banana.
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