为研究沙冬青脱水素的功能,根据沙冬青cDNA文库中脱水素基因序列设计特异引物,采用PCR技术扩增脱水素基因的DNA全长,并对其内部的一个碱基进行了定点突变,然后成功地将其连接到pCAMBIA3301载体中,用冻融法将该表达载体质粒导入根癌农杆菌菌株LBA4404中,通过农杆菌介导法,对糖用甜菜进行了遗传转化,获得了Km抗性的甜菜植株,生根后移栽到花盆中.
李晓东, 白晨, 张玉金, 于卓, 孙国琴, 张惠忠, 郭九峰, 康连和
. 沙冬青脱水素基因的克隆、重组及对糖料作物的遗传转化[J]. 华北农学报, 2010
, 25(1)
: 68
-74
.
DOI: 10.7668/hbnxb.2010.01.013
In order to study function of Ammopiptanthus mongolicus dehydrin gene,First,the primers were designed according to the sequence of Ammopiptanthus mongolicus dehydrin cDNA library,the dehydrin gene was amplified with PCR,then carried out site-directed mutagenesis for its a base internal,and cloned into pCAMBIA3301 vector.The dehydrin gene was introduced into sugar beet by Agrobacterium mediated tumerfaciens,and resistance to Km of Sugar beet were obtained,which lays a good foundation for the further research.
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