摘要
为验证虎杖白藜芦醇合酶基因PcRS在转基因植物中合成白藜芦醇的有效性,构建了植物表达载体pCAM1380-35S-PcRS,通过农杆菌介导,利用花序浸泡法转化拟南芥.转化子在含有潮霉素的培养基上经筛选后,利用PCR和Southern杂交技术检测,进一步证实PcRS基因已整合到拟南芥基因组.经过选育得到了遗传稳定的T3代纯合子转基因拟南芥株系.Northern杂交证实外源基因在转基因拟南芥纯合子株系中实现表达,并且通过HPLC法检测到终产物是反式白藜芦醇苷.两周大拟南芥幼苗产生的反式白藜芦醇苷为136μg/g(鲜重),干重为1813μg/g.
In order to reveal the effectiveness of Polygonum cuspidatum resveratrol synthase（PcRS） gene in improving the resveratrol accumulation in foreign species,first,the plant expression vector pCAM1380-35S-PcRS was constructed and introduced into Arabidopsis thaliana by Agrobacterium tumefaciens by means of the floral dip me-thod.Next,the transformants were screened on the medium containing hygromycin.Then,the integration of PcRS gene into the transgenic plants was verified via the PCR and the Southern blotting.Moreover,independent homozygous transgenic Arabidopsis thaliana lines with genetical stability were obtained after the selection of T3 progenies,and the expression of the gene transferred into Arabidopsis thaliana was confirmed by Northern blotting.Finally,an end product,namely trans-piceid,was identified by HPLC,the content of which in 2-week-old plants being 136 μg/g（fresh weight） or 1 813 μg/g（dry weight）.
出处
《华南理工大学学报（自然科学版）》
EI
CAS
CSCD
北大核心
2011年第5期138-142,共5页
Journal of South China University of Technology(Natural Science Edition)
基金
广东省自然科学基金资助项目(10151001002000012)
广东省科技计划项目(2010B060200009)
关键词
虎杖
白藜芦醇合酶
转基因
拟南芥
白藜芦醇
白藜芦醇苷
Polygonum cuspidatum
resveratrol synthase
transgene
Arabidopsis thaliana
resveratrol
piceid
