摘要
用300mmol/L NaCl处理拟南芥幼苗,分别于处理后0、1、2、4、8、16、24、48h通过Northern Blot检测其AtPUB18基因的表达量。结果显示:拟南芥AtPUB18基因的表达量受高盐胁迫的诱导而升高,于处理后4h表达量达到最高,处理后16h表达量最低。采用PCR技术克隆AtPUB18的启动子,序列为1 974bp;序列分析发现启动子内含有大量与非生物胁迫相关的顺式作用元件,如HSE、LTR、MBS及ABRE;将启动子克隆到表达载体pCambia1300-221-GUS中,驱动报告基因GUS表达。组织化学染色结果表明,未经过高盐处理的幼苗中GUS基因表达水平很低;300mmol/L NaCl处理后GUS基因表达量显著升高。研究表明,AtPUB18的表达受高盐胁迫诱导,且AtPUB18基因的启动子是一个盐胁迫诱导型启动子。
Young seedlings were treated by 300 mmol/L NaC1 and harvested after being treated for 0,1,2, 4,8,16,24 and 48 hours for RNA extraction. Northern Blot was performed to check the expression of At- PUB18. The result showed that the expression was induced by high salinity stress and reached the peak af- ter treatment for 4 h followed by decreasing to the lowest level after 16 h treatment. PCR was performed to clone the promoter of AtPUB18 which is composed of 1 974 bp. Analysis of the sequence of this promoter displayed that many cis-elements associated with abiotic stress localized in promoter, such as HSE, LTR, MBS and ABRE. The promoter were cloned into pCambia1300-221-GUS to drive the expression of GUS. Histochemical staining revealed that the expression level of GUS without salt treatment was very low, but the expression of GUS became much stronger after 4 h of 300 mmol/L NaC1 treatment. Our results mani- fested that the expression of AtPUB18 can be induced by salt stress and the promoter of AtPUB18 is a high salinity-induced promoter.
出处
《西北植物学报》
CAS
CSCD
北大核心
2014年第1期54-59,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
兵团博士资金专项(2013BB003)
石河子大学高层次人才启动项目(RCZX201218)
