AtDME1化学诱导表达载体的构建及其瞬时表达  

Constructing A Chemical-inducible Expression Vector of AtDME1 and Its Transient Expression

作　　者：常英英[1] 高亚南[1] 梁立雄 丁昌俊[1] 苏晓华[1] 张冰玉[1] 

机构地区：[1]国家林业局林木培育重点实验室(中国林业科学研究院林业研究所),北京100091

出　　处：《东北林业大学学报》2016年第2期75-79,共5页Journal of Northeast Forestry University

基　　金：林木遗传育种国家重点实验室基本科研业务费专项资金课题(TGB2013010);国家高技术研究发展计划(863计划)课题(2013AA102703)

摘　　要：以拟南芥去甲基化转移酶基因DME1为目的基因,采用酶切、连接和重组反应的方法构建了以17-β-雌二醇为诱导剂的植物表达载体p ER8-DME1。将其转入农杆菌LBA4404菌株中,通过烟草瞬时表达技术,对该载体的诱导表达特性进行了研究。q PCR结果表明,较低浓度的17-β-雌二醇处理能够有效调控p ER8-DME1中目的基因的表达,诱导处理后目的基因表达量逐渐升高,在12 h后达到最高,之后缓慢下降。该载体的成功构建为深入研究DNA去甲基化与植物的生殖发育调控奠定了基础。Dethylansferase gene DME1 from Arabidopsis thaliala was used as the target gene,and digestion,ligation and recombination reaction methods were applied for constructing the plant expression vector pE R8-DME1,which could be induced by 17-β-estradiol. The vector was transferred to Agrobacterium tumefaciens LBA4404,and the inducible expression characteristics were verified by the transient expression system of Nicotiana tabacum. By qP CR,low concentration of 17-β-estradiol could be effective for the expression of the target gene in pE R8-DME1 vector. The expression level of DME1 gene gradually increased with 17-β-estradiol treating time and reached the highest level at 12 h.

关 键 词：pER8-DME1 17-Β-雌二醇 化学诱导表达载体 瞬时表达 

分 类 号：Q782[生物学—分子生物学]