CAJ | 학술논문

启动子在基因表达调控中起关键性作用,它在很大程度上决定所控基因表达的时间、空间和强度.依据拟南芥ATH1芯片分析杨树维管形成时期特异表达基因的结果,选取了差异表达基因NST3,通过BLAST比对在杨树EST数据库(PopulusDB)中找到同源性较高的基因NAC068.以毛白杨为材料,在其基因组中克隆得到该基因5＇侧翼区901 bp长的片段,命名为pProNAC068,将该片段置换pBI121载体中的CaMV 35S启动子,并在84K杨中检测报告基因GUS的表达情况.经过GUS活性检测分析发现:该启动子可以控制外源基因在次生维管组织中特异表达,从而为基因工程中有目的的控制外源基因在维管组织中的表达奠定基础.
계동자재기인표체조공중기관건성작용,타재흔대정도상결정소공기인표체적시간、공간화강도.의거의남개ATH1심편분석양수유관형성시기특이표체기인적결과,선취료차이표체기인NST3,통과BLAST비대재양수EST수거고(PopulusDB)중조도동원성교고적기인NAC068.이모백양위재료,재기기인조중극륭득도해기인5＇측익구901 bp장적편단,명명위pProNAC068,장해편단치환pBI121재체중적CaMV 35S계동자,병재84K양중검측보고기인GUS적표체정황.경과GUS활성검측분석발현:해계동자가이공제외원기인재차생유관조직중특이표체,종이위기인공정중유목적적공제외원기인재유관조직중적표체전정기출.
Promoter plays a key role not only in regulating the level of gene expression but also in controlling the expression in a spatial and temporal manner. According to the expression profile generated using Arabidopsis ATH1 genechip to analyze genes involved in the regeneration of the secondary vascular system in Populus tomentosa Carr, a differentially expressed gene NST3 was chosen and its homologous gene NAC068 in poplar were found using BLAST software against poplar EST database (PopulusDB). Then the 5′- upstream sequence of this gene was cloned from Populus tomentosa Carr. and put into the plant expression vector pBI121 to replace the 35S promoter to control the reporter gene gus. After transformation of polar 84K via Agrobactera, the GUS activity was detected in cambium zone, suggesting this promoter could control gene expression in secondary vascular system thus should be useful in genetic engineering to control exogenous gene specific expression in vascular system.