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园艺学报 ›› 2016, Vol. 43 ›› Issue (11): 2251-2256.doi: 10.16420/j.issn.0513-353x.2016-0130

• 研究报告 • 上一篇    下一篇

菠萝AcSERK1 启动子的转录起始位点及胚性细#br# 胞特异性鉴定

栾爱萍,何业华*,林文秋,陈程杰,冯筠庭,谢 桃,龚 雪,夏靖娴   

  1. 华南农业大学园艺学院,广州 510642
  • 出版日期:2016-11-25 发布日期:2016-11-25
  • 基金资助:

    国家自然科学基金项目(30971984);公益性行业(农业)科研专项(201303021);广东省科技计划项目(2013B020304002);
    广东省现代农业产业技术体系项目(2016LM1128)

Identification of Transcription Start Site of AcSERK1 and Its Embryospecific#br# Cell Promoter in Pineapple

LUAN Ai-ping,HE Ye-hua*,LIN Wen-qiu,CHEN Cheng-jie,FENG Jun-ting,XIE Tao,GONG Xue,#br# and XIA Jing-xian   

  1. College of Horticulture,South China Agricultural University,Guangzhou 510642,China
  • Online:2016-11-25 Published:2016-11-25

摘要:

AcSERK1 是菠萝(Ananas comosus)体细胞胚发生初期特异表达的基因,为探讨其转录调控
规律,对其5′上游调控序列的转录起始位点及启动特性进行了鉴定。采用hiTAIL-PCR 技术从菠萝基因组
DNA 中克隆了AcSERK1 完整的5′上游调控序列,利用5′RACE 技术鉴定出其转录起始位点位于起始密码
子(ATG)上游258 nt(G)处。以AcSERK1 5′上游完整调控序列取代pBI121 中的CaMV 35S 启动子,构
建植物表达载体AcSERK1(–2 090/+258)︰︰GUS,并在菠萝不同组织器官中进行瞬时转化分析,发现AcSERK1
5′上游完整的调控序列(–2 090/+258)能够启动GUS 在胚性细胞中特异性表达,与AcSERK1 表达规律相
同,说明该启动子为胚性细胞特异性启动子,对调控外源基因在体细胞胚早期特异表达有重要意义。

关键词: 菠萝, AcSERK1, 启动子, 胚性细胞特异性, 转录起始位点

Abstract:

AcSERK1 is preferentially expressed during the early stage of somatic embryogenesis in
pineapple(Ananas comosus). In order to study its transcriptional regulation,the transcription start site and
starting characteristics of the 5′ upstream regulatory sequences were identified. The full length of its 5′
upstream regulatory sequence was isolated by hiTAIL-PCR from pineapple genomic DNA. The
transcription start site confirmed by 5′ RACE was located at 258 nt from 5′ upstream of the translation start
codon(ATG). A plant expression vector,AcSERK1(–2 090/+258)︰︰GUS,was constructed by inserting
AcSERK1(–2 090/+258)into the plasmid vector pBI121 whose promoter CaMV 35S was replaced.
Transient expression of GUS ananlysis of the promoter activity in different tissues and organs of pineapple
was conducted. All above demostrated that the promoter of AcSERK1 turned on GUS expression only in
embryogenic cell,which is the same as its behaviour on AcSERK1.

Key words: Ananas comosus, AcSERK1, promoter, embryo-specific cell, transcription start site

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