中国水稻科学

中国水稻科学

• 研究报告 • 上一篇    下一篇

水稻精细胞优势表达基因RSG6启动子的克隆及表达

兰利琼; 苗 琛*; 白 洁; 徐莺; 王胜华; 唐 琳; 颜钫; 陈放**   

  1. 四川大学 生命科学学院, 四川 成都 610064
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2004-07-10 发布日期:2004-07-10

Cloning and Expression of Promoter Fragment of RSG6, an Abundantly Expressed Gene in Rice Sperm Cells

LAN Li qiong; MIAO Chen*; BAI Jie; XU Ying; WANG Sheng hua; TANG Lin; YAN Fang; CHEN Fang**   

  1. College of Life Sciences; Sichuan University; Chengdu 610064; China; *Present address: College of Life Sciences; Henan University; Kaifeng 475001; China
  • Received:1900-01-01 Revised:1900-01-01 Online:2004-07-10 Published:2004-07-10

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被引次数

13

摘要： 利用已知的[i]RSG6[/i]基因序列和GenBank中提供的[i]RSG6[/i]前导序列设计引物，通过基因组DNA的PCR扩增得到长度为1 408 bp和1 173 bp的两个[i]RSG6[/i]启动子片段PB、PS，测定序列分析发现，PB、PS分别位于水稻第10和第4染色体上，PB、PS序列之间具有较高的同源性，且都含有大量的启动子元件。通过设计带有酶切位点的引物扩增出PB和PS的各两条缺失片段PB1、PB2、PS1、PS2，与质粒pBI221连接后得到中间载体pBI221PB1、pBI221PB2、pBI221PS1、pBI221PS2，再与质粒pBIN19连接构建出双元表达载体pBINGUSB1、pBINGUSB2、pBINGUSS1、pBINGUSS2，转化农杆菌EHA105，感染烟草叶片和烟草花粉，瞬时表达显示缺失片段PB1、PB2、PS1、PS2均具有启动子功能，且PB1、PS1的启动效率较PB2、PS2高。

关键词: 基因, 水稻, 启动子, 缺失片段, 瞬时表达

Abstract: Specific primers were designed according to the published genomic sequences of rice in GenBank and RSG6 gene sequence of rice sperm cells. Two 5′upstream sequence of RSG6,1 408 bp and 1 173 bp, were amplified by polymerase chain reaction(PCR) technique from DNA template of rice variety Guichao 2 and were named as PB and PS respectively. The results analyzed by NCBI BLAST and internet software PLANT CARE indicated that PB and PS are rather similar, and both fragments had many conservative promoter elements of higher plants, which suggested that PB and PS might perform their function to promote the specific expression of RSG6 in rice sperm cells. Four deletion fragments, PB1, PB2, PS1 and PS2 were produced by PCR. After entry vector pBI221 PB1, pBI221 PB2, pBI221 PS1 and pBI221 PS2 were constructed, four plant expression vectors, pBIN GUSB1, pBIN GUSB2, pBIN GUSS1 and pBIN GUSS2 were then constructed. Recombined vectors were transferred into Agrobacterium tumefaciens EHA105, and the latter were used to transform the leaves and pollen of tobacco. GUS transient expression demonstrated that the four deletion fragments could work as promoter.

Key words: gene, rice, promoter, deletion fragment, transient expression