花粉管通道法验证花生Oleosin基因启动子的特异性表达  

Identification of Specific Expression of Promoter of Oleosin Gene via Pollen-Tube Pathway Method

作　　者：范乾程[1] 谭玲玲[1] 王亚[1] 乔利仙[1] 王晶珊[1] 隋炯明[1] 

机构地区：[1]青岛农业大学生命科学学院山东省高校植物生物技术重点实验室,青岛266109

出　　处：《中国粮油学报》2013年第12期52-56,共5页Journal of the Chinese Cereals and Oils Association

基　　金：国家自然基金(31301356和31101178);山东省自然基金(ZR2012CM014,ZR2011CQ026)

摘　　要：油体蛋白(Oleosin)是一类覆盖在油体表面且仅在种子中特异表达的低分子质量蛋白,能维护油体的稳定性。研究利用花生Oleosin基因的启动子替换了pCAMBIA1301的35S启动子,构建了重组表达载体pCAMBIA1301-Oleosin-pro,并利用花粉管通道法导入花生。对T0代转基因植株进行了PCR鉴定和GUS组织化学染色,结果显示:50株转基因植株有9株能扩增出目的条带,阳性率为18%,9株阳性转基因植株中有5株的子叶能被GUS染色。对GUS染色成阳性的转基因植株的子叶进行切片观察,结果显示Oleosin基因启动子驱动的GUS基因在油体蛋白中能够表达。Oleosin is one kind of low molecular weight proteins which can be attached specifically to oil bodies to maintain the stability. In the experiment described in the paper, the recombinant expression vector pCAMBIA1301 - Oleosin - pro has been constructed by the promoter of Oleosin gene from peanut and lead - to peanut via pollen - tube pathway method. Transgenic plants of To generation were analyzed by PCR and GUS staining of tissues. The results showed that the objective band would be amplified in 9 of 50 transgenie plants with positive rate of 18% ,and GUS ac- tivity was detected only in cotyledons of 5 transgenic plants. Cotyledons of positive transgenic plants identified by GUS staining was observed by microseetion, which revealed that the GUS gene driven by the promoter of the Oleosin gene was expressed in the oil bodies.

关 键 词：花生 Oleosin基因 特异性表达 花粉管通道法 遗传转化 GUS染色 

分 类 号：Q786[生物学—分子生物学]