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拟南芥抗病相关基因T1N6_22的原核表达分析

DOI: 10.7668/hbnxb.2015.01.012, PP. 73-76

Keywords: 拟南芥,T1N6_22,原核表达,纯化

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Abstract:

为构建拟南芥抗病相关T1N6_22基因的原核表达载体并进行高效表达,获得纯化的T1N6_22蛋白,以拟南芥Col-0的cDNA为模板,扩增获得了T1N6_22基因CDS,对其进行克隆、测序后与含有GST标签蛋白的pGEX4T-1载体相连,构建T1N6_22的原核表达载体pGEX4T-1-T1N6_22-GST。经酶切和测序鉴定正确后,将构建好的载体及空载体转化大肠杆菌BL21。结果表明,在大肠杆菌BL21菌株中成功表达了与标签蛋白融合的T1N6_22蛋白,大小约57kDa。SDS-PAGE分析表明,高效表达的诱导条件为0.1mmol/LIPTG诱导培养3h。研究结果为进一步T1N6_22互作蛋白的筛选及其调控拟南芥抗病分子机制的研究奠定了基础。

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