番木瓜proteinase omega 基因启动子的克隆及功能初步研究

Plant Diversity ›› 2005, Vol. 27 ›› Issue (05): 545-551.

番木瓜proteinase omega 基因启动子的克隆及功能初步研究

杨英军^{1 , 2} , 周鹏¹

1 中国热带农业科学院热带生物技术研究所; 2 河南科技大学

收稿日期:2005-03-11 出版日期:2005-10-01 发布日期:2005-10-01
通讯作者: 杨英军

Cloning and Functional Analysis of a Novel Proteinase Omega Promoter from Carica papaya

YANG Ying-Jun^{1 , 2}, ZHOU Peng¹

1 National Key Biotechnology Laboratory for Tropical Crops, Institute of Bioscience and Biotechnology , CATAS , Haikou , 571101 , China ; 2 Henan University of Science and Technology , Luoyang 471003 , China

Received:2005-03-11 Online:2005-10-01 Published:2005-10-01
Contact: YANG Ying-Jun

摘要/Abstract

摘要： 采用PCR 技术从番木瓜基因组中克隆了proteinase omega 基因的部分序列及其5′侧翼序列。序列分析表明, 克隆到的基因序列与GenBank 中的序列同源性为96%, 长1039 bp 的5′端侧翼序列在GenBank 数据库中没有同源片段。预测5′端侧翼序列有两处基础启动子区域,转录起始位点(TSS) 分别为是A, T。在基础启动子区域都存在TATA-box, 上游发现多处CAAT-box , G-box , I-box 等顺式作用元件和AT 富含区。构建了植物表达载体并用基因枪轰击番木瓜的叶组织, GUS 基因瞬间表达结果表明, 该长1 039 bp 的5′端侧翼序列具有驱动GUS基因在乳管中表达的功能。该启动子的发现对进一步研究启动子的功能和开发番木瓜作为生物反应器具有重要意义。

关键词: 番木瓜, proteinase omega, , 启动子, 克隆, 序列分析FONT

Abstract: Proteinase omega was a kind of proteinase in papaya (Carica papaya L .) laticifers . A 5′flanking sequence and partial gene sequence of proteinase omega were isolated from the genomic DNA via PCR technology . DNA sequence analysis and homology comparison indicated that it had 96% homology with the proteinase omega gene which had been submitted to GenBank . The two core promoter regions and some upstream regulatory elements in the fragment were analysed using the software of PROMOTER PREDICTION and PLANT CARE . Transcriptional start sites ( TSS) were A, T respectively . TATA-box , CAAT-box , G-box, I-box, AT rich regions and other cis- elements were found in the promoter in identical positions common to all promoter sequence regions . Compared with the data in GenBank , the results showed that a new promoter was obtained , and its GenBank accession number was AY695444 . Binary vectors were constructed through fusing 1039 bp 5′flanking region of proteinase omega gene with the GUS
gene, Transient GUS expressions were observed in papaya leaves transferred via particle bombardment . GUS activities were detected only in laticiferious cells .

Key words: Carica papaya

杨英军1 , 2 , 周鹏1 . 番木瓜proteinase omega 基因启动子的克隆及功能初步研究[J]. Plant Diversity, 2005, 27(05): 545-551.

YANG Ying-Jun^,, ZHOU Peng. Cloning and Functional Analysis of a Novel Proteinase Omega Promoter from Carica papaya[J]. Plant Diversity, 2005, 27(05): 545-551.

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