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中国农学通报 ›› 2015, Vol. 31 ›› Issue (30): 220-224.doi: 10.11924/j.issn.1000-6850.casb15060135

所属专题: 生物技术 水稻

• 生物技术科学 • 上一篇    下一篇

水稻稻瘟病抗病基因Pi63顺式作用元件的载体构建

梁华兵,王晓婉,胡晓岚,赵 峰,张成国,韩淑慧,姚富城,徐 鑫   

  1. (中南民族大学生命科学学院/武陵山区特色资源植物种质保护与利用湖北省重点实验室/生物技术国家民委重点实验室,武汉 430074)
  • 收稿日期:2015-06-26 修回日期:2015-07-16 接受日期:2015-07-21 出版日期:2015-10-29 发布日期:2015-10-29
  • 通讯作者: 徐鑫
  • 基金资助:
    湖北省自然科学基金面上项目“稻瘟病抗性新品种的培育”(2013CFB453);中央高校专项基金“三系杂交水稻种子分子检测技术及应用”(CZZ12006)。

Vector Construction of Cis-acting Regulatory DNA Elements of Rice Blast Resistance Gene Pi63

Liang Huabing, Wang Xiaowan, Hu Xiaolan, Zhao Feng, Zhang Chengguo, Han Shuhui, Yao Fucheng, Xu Xin   

  1. (Hubei Provincial Key Laboratory for Protection and Application of Special Plants in Wuling Area of China/Key Lab for Biotechnology of State Ethnic Affairs Commission/College of Life Science, South-Central University for Nationalities, Wuhan 430074)
  • Received:2015-06-26 Revised:2015-07-16 Accepted:2015-07-21 Online:2015-10-29 Published:2015-10-29

摘要: 水稻稻瘟病抗性基因Pi63具有良好的田间抗性,且抗性与其表达量成正相关,但Pi63表达的调控机制尚未得到研究。为了研究Pi63的抗性机理,通过对Pi63启动子区域顺式作用元件的生物信息学分析,分段构建了4个含有不同顺式作用元件的缺失体,将上述缺失体转入植物双元表达载体pCAMBIA1391Z中。菌落PCR以及质粒DNA酶切鉴定后,经测序进一步确认。结果表明:以上4个缺失体载体构建成功。本研究为进一步研究启动子中不同顺式作用元件对于Pi63基因稻瘟病抗性的影响以及Pi63基因的表达特性奠定了基础。

关键词: 主成分分析, 主成分分析, GIS空间分析, 区位商, 综合评价, 关天经济区

Abstract: Rice blast resistance gene Pi63 has fine field resistance to the rice blast disease and its resistance levels are closely correlated with its gene expression levels. However, the mechanism of the expression of Pi63 has not been investigated. In this study, cis-acting regulatory DNA elements in promoter region of Pi63 were predicted by bioinformatics analysis. 4 constructs containing different elements were obtained using plant binary vector pCAMBIA1391Z. The positive clones were verified by colony PCR and restriction enzyme digestion and further confirmed by Sanger sequencing. The results showed that the 4 deletant vectors were successfully constructed, which laid the foundation for further research on the effect of different cis-acting regulatory DNA element on the resistance of Pi63 and the expression pattern of Pi63.

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