黑果枸杞及其白色果实F3′5′H基因启动子克隆及活性分析

发布时间：2018-07-15 22:46

【摘要】：通过3次染色体步移克隆出黑果枸杞及其白化果实F3′5′H基因的启动子片段并测序。分析发现2个启动子序列的同源性高达90.3%,对启动子序列进行预测,发现两者中均具有TATA-Box、CAAT-Box、TCrich repeats、WUN-motif、Sp1、Box I、G-box、skin-1motif及ARE元件。另外在黑果枸杞启动子中预测到与茉莉酸甲酯响应相关的元件TGACG-motif,而在黑果枸杞白化果实启动子中没有预测到。将2个启动子片段与GUS报告基因融合构建植物表达载体,利用农杆菌介导的瞬时转化法转化烟草叶片,通过组织化学染色来确定启动子的启动活性。结果表明2个启动子均具有启动活性,利用荧光定量对2个启动子所驱动的GUS基因的表达量进行了分析,结果表明黑果枸杞启动子所驱动的GUS基因的表达量是黑果枸杞白化果实的3.09倍。
[Abstract]:The promoter fragment of the F3O5H gene of Lycium barbarum and its albino fruit was cloned and sequenced by three chromosome steps. It was found that the homology of the two promoter sequences was as high as 90.3, and the promoter sequences were predicted. It was found that both of them had TATA-BoxCX CAAT-BoxCrich repeatsWUN-motif Sp1BOX G-boxskin-1motif and are elements. In addition, the element TGACG-motif related to the response of methyl jasmonate was predicted in the promoter of Lycium barbarum, but not in the promoter of albino fruit of Lycium barbarum L. The plant expression vector was constructed by fusion of two promoter fragments with Gus reporter gene and transformed into tobacco leaves by Agrobacterium tumefaciens-mediated transient transformation. The promoter activity was determined by histochemical staining. The results showed that both promoters had priming activity. The expression of Gus gene driven by two promoters was analyzed by fluorescence quantitative analysis. The results showed that the expression of Gus gene driven by the promoter of Lycium barbarum was 3.09 times higher than that of the albino fruit of Lycium barbarum L.
【作者单位】：青海大学农林科学院青海高原林木遗传育种实验室;青海大学省部共建三江源生态与高原农牧业国家重点实验室;
【基金】：国家自然科学基金地区科学基金项目(31660221) 青海省农林科学院创新基金(2014-NKY-01) 青海高原林木遗传育种重点实验室(2017-ZJ-Y29)
【分类号】：S567.19

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