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CMS结球甘蓝剪接异构体BoKIN1-2的鉴定及其表达研究

DOI: 10.11869/j.issn.100-8551.2015.11.2077, PP. 2077-2083

Keywords: CMS结球甘蓝,KIN基因,BoKIN1-2,选择性剪接,基因表达

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Abstract:

为了探讨结球甘蓝BoKIN1基因的pre-mRNA加工机制的特点,以细胞质雄性不育系(CMS)结球甘蓝为试验材料,使用特异引物进行RT-PCR和荧光定量PCR,分析抗冻蛋白BoKIN1基因的剪接异构体BoKIN1-2的特征特性。结果显示,BoKIN1-2的PCR产物溶解曲线的Tm值为84.41℃。在第2个外显子和第2个内含子的连接处设计引物进行3'RACE分析,测序结果显示BoKIN1-2包含第2个内含子和ploy(A)尾,证明BoKIN1基因在pre-mRNA加工过程中保留了第2个内含子。荧光定量PCR结果显示,BoKIN1-2在茎、叶、花、雄蕊和雌蕊中均有表达,在高温、低温和ABA处理后的叶片中表达上升。BoKIN1基因的选择性剪接既参与了器官发育也参与了胁迫响应。本研究为后期分析KIN基因的转录后调控分子机制提供了科学依据。

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