摘要
采用RT-PCR法克隆了蒺藜苜蓿MtCKX基因,基因编码区长1635bp,编码544个氨基酸。生物信息学分析表明,其编码的蛋白与其他物种CKX蛋白具有较高的同源性,与鹰嘴豆的同源性最高。荧光定量分析结果表明,MtCKX基因在蒺藜苜蓿叶片中的表达量最高。此外,MtCKX基因还响应IAA、6-BA的诱导。成功构建了35S::MtCKX:YFP植物表达载体,亚细胞定位分析表明,MtCKX定位于细胞膜和细胞核。过表达的蒺藜苜蓿MtCKX基因可导致转基因拟南芥中细胞分裂素含量显著降低。
CKX,a key factor that manipulates plant endogenous cytokinin content,is considered as the target gene that regulates crop yield and resistance.MtCKX was cloned by RT-PCR from Medicago truncatula.The coding domain sequence was 1635 bp in length,encoding 544 amino acids.Based on bioinformatic analysis,MtCKX showed highly homologous characteristics with other CKX family genes,especially homologous with CKX from Cicer anetinum.Quantitative Real-time PCR showed that the expression of MtCKX was highest in the leaf blade.MtCKX was responsive to the induction of IAA and 6-BA.CKX gene was inserted into 3302 Yplasmid to construct the plant expression vector 35 S：：MtCKX：YFP.Subcellular localization analysis showed that MtCKX localized in the membrane and nucleus.Over expression of MtCKX gene resulted in a significant decrease in cytokinin content in transgenic Arabidopsis thaliana.
作者
李殷睿智
韩东英
刘卓成
滕珂
晁跃辉
韩烈保
LI Yin-rui-zhi;HAN Dong-ying;LIU Zhuo-cheng;TENG Ke;CHAO Yue-hui;HAN Lie-bao(Turfgrass Research Institute, Beijing Forestry University, Beijing 100083, China)
出处
《中国草地学报》
CSCD
北大核心
2018年第2期1-7,共7页
Chinese Journal of Grassland
基金
国家自然基金项目青年科学基金项目(31601989)
深圳市科技计划项目(JCYJ20160331151245672
JSGG20160229155434792)
中国博士后基金(2017M620677)
北京市博士后基金(2017-ZZ-087)
关键词
蒺藜苜蓿
MtCKX基因
亚细胞定位
表达分析
Medicago truncatula
MtCKX
Subcellular localization
Quantitative Real-time PCR
