| 摘要: |
| 根据拟南芥PIN3 cDNA序列设计引物,通过逆转录PCR(Reverse transcription-polymerase chain reaction)克隆荠菜中PIN3的同源性cDNA。同时利用染色体步移法从荠菜基因组中克隆该基因上游1 488 bp含启动子的基因组DNA序列。序列分析表明,荠菜PIN3基因cDNA全长1 950 bp,可编码1个含有649个氨基酸的推导蛋白。荠菜PIN3 cDNA与拟南芥PIN3 cDNA具有91%的同源性,2种PIN3推导蛋白的同源性也高于90%。将克隆的荠菜PIN3基因启动子序列与拟南芥同源基因启动子序列进行比较分析,两者同源性为82%。荠菜PIN3启动子含有真核生物典型的TATA-BOX、CAAT-BOX、光响应元件、水杨酸响应元件、厌氧诱导响应元件和胚乳高水平表达响应元件等。与拟南芥PIN3启动子相比,二者除了含有相同的启动子基本元件和部分光响应元件外,还含有差异性的光响应元件以及不同的与胁迫反应相关的顺式作用元件。判断上述差异可使二者在心皮发育过程中通过不同的生长素运输模式调控其形态发育。 |
| 关键词: 荠菜 PIN3基因 PIN3启动子 克隆 序列分析 |
| DOI：10.11841/j.issn.1007-4333.2015.01.004 |
| 投稿时间：2014-04-03 |
| 基金项目:湖南省科技计划项目(2012NK3062) |
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| Cloning and characterization of auxin polar transport PIN3 gene cDNA and its promoter in Capsella bursa-pastoris |
|
HU Qing-yun, ZHU Zhan-wei, PENG Yan, ZHAO Yan, HUANG Yu, LIU Xiao-zhu, ZHANG Xue-wen
|
| (College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China) |
| Abstract: |
| In this paper the primers were designed according to the PIN3 gene cDNA of Arabidopsis.Homologous gene of PIN3 in Capsella bursa-pastoris was cloned by RT-PCR(Reverse transcription-polymerase chain reaction).A 1 488 bp upstream fragment of PIN3 gene was also cloned from Capsella bursa-pastoris genome by chromosome walking method.Sequence analysis showed that the cDNA was 1 950 bp in length and encoded a putative protein of 649 amino acids.PIN3 cDNA in Arabidopsis and Capsella bursa-patoris had 91% homology and their putative protein homologous was higher than 90%.The promoter of PIN3 gene of Capsella bursa-patoris shared 82% homology with its counterpart in Arabidopsis.The PIN3 gene promoter in Capsella bursa-passtoris had the TATA-BOX and CAAT-BOX which was Eukaryotes typical elements and some other regulatory elements like light responsive element,cis-acting regulatory element involved in the salicylic acid responsiveness,cis-acting regulatory element involved in anaerobic induction,cis-acting regulatory element involved in high level expression of endosperm that were found in the Arabidopsis.But there were some different light responsive elements and a few cis-acting regulatory elements involved in abiotic stress response specifically contained in the Capsella bursa-patoris promoter.The difference elements may lead to a different auxin transportation model in the carpel and build a different carpel morphology. |
| Key words: Capsella bursa-pastoris PIN3 gene PIN3 promoter cloning sequence analysis |
