摘要
在前期的研究中,一个玫瑰RdGASA4-like基因的cDNA和DNA序列全长利用同源克隆结合RACE的方法首次得到克隆。为深入了解该基因在赤霉素调控植物发育过程中的作用,通过TAIL-PCR的方法首次克隆该基因起始密码子ATG上游685bp的5’-UTR和部分启动子序列,在启动子序列分析的基础上,构建其与GUS基因融合表达载体,命名为pBI-GP685。通过农杆菌介导的方法对甘蓝外植体子叶-子叶柄进行启动子瞬时表达研究,结果表明分离得到的启动子具有启动报告基因GUS表达的活性。
In previous study,the full length DNA and cDNA of theRdGASA 4-like gene had been isolated from Damask rose(Rosa damascena )through homology cloning combined with rapid amplification of cDNA ends (RACE).In order to further understand the role ofRdGASA 4-like in the regulating network on plant development of gibberellic acid,its 685 bp 5’-UTR and partial sequence upstream ATG ofRdGASA 4-like was obtained by Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL-PCR).Based on sequence analysis,it was fused to a GUS gene to construct plant expression vector,named as pBI-GP685.By agrobacterium-mediated method,the explants of petiole with cotyledon of cabbage were transformed.The transient expression study showed that the obtained promoter ofRdGASA 4-like was active and could drive the activity of GUS.
出处
《中国农学通报》
CSCD
北大核心
2011年第8期123-127,共5页
Chinese Agricultural Science Bulletin
基金
河南省基础与前沿技术研究计划项目"玫瑰花香基因克隆及其对月季的遗传转化"(082300430100)