苦荞锌指蛋白基因FtLOL1的克隆及其对非生物胁迫的应答  

Cloning and Expression Analysis One Zinc Finger Protein Gene Ft LOL1 in Fagopyrum tataricum: Effect of Abiotic Stress

作　　者：高飞[1] 周婧[1] 刘晓彤[1] 李成磊[1] 姚慧鹏[1] 赵海霞[1] 吴琦[1] 

机构地区：[1]四川农业大学生命科学院,雅安625014

出　　处：《中国生物工程杂志》2015年第8期44-50,共7页China Biotechnology

基　　金：四川省教育厅青年基金资助项目(13ZB0294)

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Ft LOL1(Ft LSD-one-like 1,Gen Bank登录号:KP260662),获得c DNA和DNA全长序列,采用实时荧光定量PCR技术研究Ft LOL1在3种非生物处理条件下的表达模式。结果显示,苦荞Ft LOL1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;c DNA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Ft LOL1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,Ft LOL1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。According to transcriptome data of Fagopyrum tataricum at flowering,using PCR and RT-PCR techniques,the DNA and c DNA sequences of Ft LOL1（ Gen Bank accession number： KP260662） gene were amplified from Fagopyrum tataricum. The obtained sequences were analyzed by bioinformatics software,and the expression Ft LOL1 gene were analysed by q PCR under UV-B,SA and 4℃ cold stress. The results showed that the DNA sequence of Ft LOL1 gene was 1 720 bp,of which consisted 5 exons and 4 introns,in line with the principle of GT-AG splicing,and the c DNA of Ft LOL1 contained a 444 bp 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 Ft LOL1 gene. The results expected to lay a foundation for the stress resistance study in Fagopyrum tataricum.

关 键 词：苦荞 锌指蛋白 基因克隆 非生物胁迫 表达 

分 类 号：Q789[生物学—分子生物学]