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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2015, Vol. 35 Issue (8): 44-50    DOI: 10.13523/j.cb.20150807
研究报告     
苦荞锌指蛋白基因FtLOL1的克隆及其对非生物胁迫的应答
高飞, 周婧, 刘晓彤, 李成磊, 姚慧鹏, 赵海霞, 吴琦
四川农业大学生命科学院 雅安 625014
Cloning and Expression Analysis One Zinc Finger Protein Gene FtLOL1 in Fagopyrum tataricum: Effect of Abiotic Stress
GAO Fei, ZHOU Jing, LIU Xiao-tong, LI Cheng-lei, YAO Hui-peng, ZHAO Hai-xia, WU Qi
College of Life Science, Sichuan Agricultural University, Ya'an 625014, China
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摘要:

根据苦荞(Fagopyrum tataricum)花期转录组数据,采用RT-PCR技术和PCR技术克隆得到一个C2C2型锌指蛋白基因FtLOL1(FtLSD-one-like 1,GenBank登录号:KP260662),获得 cDNA和DNA全长序列,采用实时荧光定量PCR技术研究FtLOL1在3种非生物处理条件下的表达模式。结果显示,苦荞FtLOL1基因DNA全长1 720bp,由5个外显子和4个内含子组成,符合GT-AG剪切原则;cDNA包含一个444 bp的开放阅读框,编码147个氨基酸,具有LSD1家族的典型特征;2mmol/L水杨酸、UV-B照射和4℃冷处理均能导致FtLOL1基因表达量下降,其中水杨酸和UV-B处理均在12h达到最小值,分别为对照组(0h)的11.9%和33.8%,而4℃冷处理条件下,FtLOL1表达量12h开始下降,至24h达到最小值,为对照组(0h)的50.7%,36h后表达量有所回升,稳定在对照组(0h)的60%左右。推测该基因可能参与苦荞抗高浓度水杨酸、UV-B照射和冷胁迫等非生物胁迫的应答反应,为探究苦荞抗逆性提供了新的视角。

关键词: 苦荞锌指蛋白基因克隆非生物胁迫表达    
Abstract:

According to transcriptome data of Fagopyrum tataricum at flowering, using PCR and RT-PCR techniques, the DNA and cDNA sequences of FtLOL1 (GenBank accession number: KP260662) gene were amplified from Fagopyrum tataricum. The obtained sequences were analyzed by bioinformatics software, and the expression FtLOL1 gene were analysed by qPCR under UV-B, SA and 4℃ cold stress. The results showed that the DNA sequence of FtLOL1 gene was 1 720bp, of which consisted 5 exons and 4 introns, in line with the principle of GT-AG splicing, and the cDNA of FtLOL1 contained a 444bp ORF, encoding 147 amino acids, with the typical features of LSD1 family. The 2mmol/L SA, UV-B radiation and 4℃ cold stress could lead to a significant discrease in the expression of the FtLOL1 gene. The results expected to lay a foundation for the stress resistance study in Fagopyrum tataricum.

Key words: Fagopyrum tataricum    Zinc finger protein LSD1    Gene cloning    Abiotic stress    Gene expression
收稿日期: 2015-03-11 出版日期: 2015-08-25
ZTFLH:  Q789  
基金资助:

四川省教育厅青年基金资助项目(13ZB0294)

通讯作者: 赵海霞     E-mail: aasky8@163.com
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引用本文:

高飞, 周婧, 刘晓彤, 李成磊, 姚慧鹏, 赵海霞, 吴琦. 苦荞锌指蛋白基因FtLOL1的克隆及其对非生物胁迫的应答[J]. 中国生物工程杂志, 2015, 35(8): 44-50.

GAO Fei, ZHOU Jing, LIU Xiao-tong, LI Cheng-lei, YAO Hui-peng, ZHAO Hai-xia, WU Qi . Cloning and Expression Analysis One Zinc Finger Protein Gene FtLOL1 in Fagopyrum tataricum: Effect of Abiotic Stress. China Biotechnology, 2015, 35(8): 44-50.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150807        https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I8/44


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