雪莲SikCDPK1启动子的克隆和活性分析  

Cloning and Activity Analysis of SikCDPK1 Promoter from Saussurea involucrata

作　　者：史光珍 王兆晔 孙琦 朱新霞[1] SHI Guang-zhen;WANG Zhao-ye;SUN Qi;ZHU Xin-xia(School of Life Sciences,Shihezi University,Key Laboratory of Oasis Town and Mountain Basin System Ecology Corps,Shihezi 832003)

机构地区：[1]石河子大学生命科学学院绿洲城镇与山盆系统生态兵团重点实验室,石河子832003

出　　处：《生物技术通报》2022年第9期191-197,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31760066)。

摘　　要：旨在克隆雪莲(Saussurea involucrata)SikCDPK1 启动子并分析其活性,为进一步解析雪莲 SikCDPK1基因的转录调控机制奠定基础。通过TAIL-PCR技术从雪莲中克隆SikCDPK1启动子,利用PlantCARE分析启动子区顺式作用元件,构建全长启动子或5’端缺失启动子驱动的GUS重组表达载体 P0∷GUS、P1∷GUS、P2∷GUS和P3∷GUS,转入根癌农杆菌中进行瞬时转化试验,通过GUS组织化学染色分析不同长度启动子的活性,分别测定低温和干旱胁迫下的GUS酶活。结果显示,获得了1 042 bp的SikCDPK1启动子序列,pSikCDPK1具有真核生物启动子核心元件TATA-box和CAAT-box,还含有多个与逆境、激素、光响应等相关的顺式作用元件。转化的烟草叶片经GUS染色之后均显蓝色,启动活性依次为P0>P1>P2>P3,低温、干旱处理后GUS酶活性发生变化。SikCDPK1启动子被成功克隆,具有驱动下游报告基因表达的活性。The objective is to clone the SikCDPK1 promoter of Saussurea involucrata and analyze its activity,so as to lay the foundation for further understanding the transcriptional regulation mechanism of the SikCDPK1 gene in S.involucrata.TAIL-PCR was used to clone the promoter of SikCDPK1 from S.involucrata,and PlantCARE to analyze the promoter cis-acting elements.The recombinant expression vector P0∷GUS,P1∷GUS,P2∷GUS and P3∷GUS driven by full-length promoter or 5’-deletion promoter were constructed and transferred into Agrobacterium tumefaciens for transient transformation.The activities of different length promoters were analyzed by GUS staining,and the activity of GUS under low temperature and drought was determined respectively.As results,a 1 042 bp promoter sequence of SikCDPK1 was obtained.pSikCDPK1 had the core elements of eukaryotic promoter TATA-box and CAAT-box,and also contained several cis-acting elements related to stress,hormone and light response.All transformed tobacco leaves showed blue color after GUS staining,and the activity was in P0>P1>P2>P3.The GUS activity changed after low temperature and drought.In sum,the promoter of SikCDPK1 is successfully cloned and has the activity of driving downstream reporter gene expression.

关 键 词：雪莲 SikCDPK1启动子 GUS活性 瞬时表达 

分 类 号：Q943.2[生物学—植物学]