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园艺学报 ›› 2013, Vol. 40 ›› Issue (5): 887-.

• 蔬菜 • 上一篇    下一篇

甘蓝Ogura 细胞质雄性不育相关基因BoMF1启动子的克隆及功能分析

 郭盈盈, 颉建明, 简元才, 郁继华, 康俊根   

  1. 1 甘肃农业大学农学院,兰州 730070;2 北京市农林科学院蔬菜研究中心,农业部华北地区园艺作物生物学与种质
    创制重点实验室,北京 100097
  • 出版日期:2013-05-25 发布日期:2013-05-25

Cloning and Functional Analysis of OguCMS-related gene BoMF1 Promoter in Brassica oleracea

 GUO  Ying-Ying, JIE  Jian-Ming, JIAN  Yuan-Cai, YU  Ji-Hua, KANG  Jun-Gen   

  1. 1Gansu Agriculture University,Lanzhou 730070,China;2Beijing Vegetable Research Center,Beijing Academy of
    Agriculture and Forestry Sciences,Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(North
    China),Ministry of Agriculture,Beijing 100097,China
  • Online:2013-05-25 Published:2013-05-25

摘要: 通过TAIL-PCR 染色体步移技术,从甘蓝(Brassica oleracea L. var. capitata L.)基因组中克
隆到胞质雄性不育(OguCMS)相关基因BoMF1 翻译起始位点上游521 bp 的启动子序列。软件分析预测
表明,该启动子序列中存在多个顺式作用元件,包括TATA-box、CAAT-box、MYB 结合位点、植物激
素响应单元等。为了研究该启动子的表达特性,亚克隆了BoMF1 转录起始位点上游521 bp 序列,将其置
换pBI121 中的CaMV35S 启动子,驱动其下游的GUS 基因,构建植物表达载体pBI121-BoMF1P,以pBI121
空载体作为阳性对照,通过农杆菌(LBA4404)介导法转入拟南芥。结果表明,甘蓝BoMF1 启动子序列
能驱动GUS 基因在拟南芥花药发育晚期的花药和花粉中特异表达,表达具有组织特异性。

关键词: 甘蓝, OguCMS, BoMF1 启动子, GUS, 调控元件

Abstract: The regulative sequence(521 bp)of OguCMS-related gene BoMF1 promoter from
Brassica oleracea was cloned by genomic walking(TAIL-PCR). In silico analysis showed that this
sequence contained several acting elements,including TATA-box and CAAT-box,MYB binding sites,
phytohormone responsive elements and so on. In order to study the promoter function,a 521 bp promoter
sequence of BoMF1 was inserted upstream of the GUS reporter gene replacing the CaMV35S promoter of
pBI121. The plant expression vector pBI121-BoMF1P and pBI121 were transformed into Arabidopsis
thaliana
with the Agrobacterium tumefaciens strain LBA4404. It was suggested that pBI121-BoMF1P
could drive the GUS gene exclusively express in anther and pollen of Arabidopsis thaliana.

Key words: Brassica oleracea, OguCMS, BoMF1 promoter, GUS, cis-element

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