园艺学报 ›› 2008, Vol. 35 ›› Issue (7): 973-978.

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杨英军1,2;周 鹏2*;李艳梅1;沈文涛2   

  1. 1 河南科技大学林学院,河南 洛阳 471003;2中国热带农业科学院热带生物技术研究所,热带作物生物技术国家重点实验室,海口571101)
  • 收稿日期:2008-01-02 修回日期:2008-05-26 出版日期:2008-07-25 发布日期:2008-07-25
  • 通讯作者: 周 鹏

Cloning and Functional Analysis of a Novel Chymopapain Promoter from Carica papaya

YANG Ying-jun1,2,ZHOU Peng2*,LI Yan-mei1, and SHEN Wen-tao2   

  1. (1College of Forestry,Henan University of Science and Technology, Luoyang Henan 471003,China; 2 National Key Biotechnology Laboratory for Tropical Crops Institute of Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou Hainan 571101,China)
  • Received:2008-01-02 Revised:2008-05-26 Online:2008-07-25 Published:2008-07-25
  • Contact: ZHOU Peng

摘要: 采用 PCR技术从番木瓜品种‘穗中红’克隆了木瓜凝乳蛋白酶基因的部分序列及其5'侧翼序列,构建含有两种长度启动子片段的植物表达载体,并用基因枪轰击番木瓜叶组织和农杆菌转化烟草叶盘。结果表明,克隆产物长719 bp,163 bp 3'侧翼序列与 GenBank中的番木瓜凝乳蛋白酶基因序列同源性为99%,556 bp的 5'侧翼序列有基础启动子区,转录起始位点位于ATG上游38bp的T。序列登录号为 AY803756。预测在启动子区存在 TATA-box、CAAT-box、WUN和HSE等顺式作用元件。两种启动子片段驱动的GUS基因瞬时表达和稳定表达结果表明,该启动子片段具有乳管特异表达活性。

关键词: 番木瓜, 木瓜凝乳蛋白酶, 启动子, 序列分析, 基因表达, 乳管


A partial gene sequence and 5' flanking sequence of chymopapain were isolated from the genomic DNA of Carica papaya via PCR technology. The results of alignment indicated that the isolated DNA sequence had 99% homology with that of the chymopapain gene in papaya. The core promoter regions and some upstream regulatory elements in this fragment were analyzed. Transcriptional start site(TSS)was T predicted by the software of PROMOTER PREDICTION and PLANT CARE TATA-box,CAAT-box,WUN,HSE regions and other cis-elements were found incooespanding promoter sequence regions with others. Compared with the data in GenBank,the results showed that a novel promoter was obtained,the GenBank accession number was AY803756. Binary vectors were then constructed,GUS expressions were both observed in papaya leaves transferred via particle bombardment and tobacco plantlet mediated by agrobacterium. GUS activities were detected only in latex.

Key words: Carica papaya, chymopapain, promoter, sequence analysis, gene expression, laticifer