马铃薯蛋白激酶基因StPK1启动子的克隆及活性分析

 【目的】研究马铃薯蛋白激酶基因StPK1,为在马铃薯抗病反应中的功能提供证据。【方法】利用 TAIL-PCR技术,从马铃薯晚疫病水平抗性、垂直抗性和感病等3种材料中,分别克隆了该基因的启动子区域。此外,将从水平抗性材料中克隆得到的启动子与GUS连接构建植物表达载体,通过农杆菌介导的遗传转化法转化本氏烟(Nicotiana benthamiana),将得到的转基因烟草分别用水、晚疫病原菌或水杨酸(SA)溶液处理,并进行表达活性分析。【结果】3个启动子长度分别为922、929和922 bp,均具有TATA-box和CAAT-box以及多种与抗病反应和基因时空表达有关的元件,而三者的主要差异是个别碱基的不同。GUS组织化学染色表明,用晚疫病原菌或SA处理的转基因烟草被染成蓝色。【结论】StPK1启动子为有活性的启动子,且为病原和SA诱导型启动子。

Cloning and Activity Analysis of the Promoter of Potato Protein Kinase Gene StPK1

【Objective】 The objective of the experiment is to study the function of potato (Solanum tuberosum L.) protein kinase gene StPK1. 【Method】 The promoters of StPK1 were cloned by TAIL-PCR from leaves of three potato cultivars with distinct resistance types to late blight: quantitative, qualitative and susceptible. The promoter cloned from the qualitative resistance potato was fused to the GUS to construct the plant expression vector, and then the chimeric gene was transferred into Nicotiana benthamiana by Agrobacterium tumefaciens-mediated method. The transformed plants were treated with water, Phytophthora infestans and salicylic acid (SA), and the expression activity of the promoter was analyzed as well. 【Result】 The length of the promoters obtained are 922 bp, 929 bp, and 922 bp, respectively. The structure comparison indicated that all the promoters possess TATA-box, CAAT-box and the elements responsive to plant disease infection as well as those related to temporal and spatial expression. Their main differences were only observed in a little change of bases. And the results demonstrated that GUS was expressed in the leaves of transgenic tobacco treated with P. infestans and SA. 【Conclusion】 It showed that the promoter is active and pathogen- and SA-inducible.