雨生红球藻多拷贝基因bkts差异表达的转录调控机理

项目名称： 雨生红球藻多拷贝基因bkts差异表达的转录调控机理

项目编号： No.31470389

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 王潮岗

作者单位： 深圳大学

项目金额： 80万元

中文摘要： β-胡萝卜素酮化酶基因（bkts）是雨生红球藻虾青素合成的关键酶基因，同时也是多拷贝基因，它们如何采取差异化转录调控来应对环境胁迫的机理还不清楚。虾青素合成的调控发生在转录水平，因此，bkts转录调控的机理也是揭示虾青素合成调控机制的关键。然而，由于雨生红球藻研究方法的滞后以及bkts启动子研究的不足，至今还没弄清虾青素合成调控的分子机制。我们前期研究克隆了bkt1和bkt2的启动子并证明了它们的功能，发现bkts各拷贝基因的启动子虽然差异很大，但都含有大量与光照和盐胁迫相关的顺式作用元件。本项目拟在前期研究基础上，以雨生红球藻和莱茵衣藻为材料，利用酵母单杂交等技术和转录组学研究筛选并鉴定出参与bkts转录调控的转录因子，再通过酵母双杂交、基因敲除和基因过表达等技术对它们的功能进行系统研究，最终阐明雨生红球藻bkts转录调控的分子机理，为利用人工调控并提高虾青素产量提供新的途径。

中文关键词： 雨生红球藻；莱茵衣藻；调控机理；β-胡萝卜素酮化酶基因；转录因子

英文摘要： β -carotene ketolase gene (bkts) known as multicopy genes is a key enzyme gene for astaxanthin biosynthesis in Haematococcus pluvialis. It is still unclear about these multicopy genes how to deal with environmental stresses with regulation in transcription differentiation in Haematococcus pluvialis. Regulation of bkts transcription might play a key role in the astaxanthin biosynthesis because references confirmed that regulation of astaxanthin biosynthesis is controlled in transcriptional level. However, we still unknow regulatory mechanism of astaxanthin biosynthesis due to a long period of stagnation in study methods and shortage of the knowledge on bkts promoter in Haematococcus pluvialis. In our previous study, both bkt1 and bkt2 promoter were cloned from Haematococcus pluvialis genomic library and their functions were confirmed by molecular methods. Sequence analysis and data search revealed that there were a lot of cis-elements associated with light and salt stress-related responses though these two promoters existed with low identity. Here, transcription factors involving in regulating bkts transcription will be screened from Haematococcus pluvialis and Chlamydomonas reinhardtii by Yeast one-hybrid system, transcriptome analyse, etc. In order to analyze the interactions and functions of these transcription factors involving in regulation of bkts transcription, some methods including Yeast two -hybrid system, gene knock-out, gene over-expression will be employed. In conclusion, regulatory mechanism of bkts transcription will be illuminated based on our study results. It will provide a new way for increasing production of astaxanthin by artificial control.

英文关键词： Haematococcus pluvialis;Chlamydomonas reinhardtii;regulatory mechanism;bkts;transcription factor