摘要
载体的构建是建立遗传转化体系的基础。以真核表达载体pcDNA3.1(-)/hygro为基本骨架,构建大豆疫霉菌遗传转化载体,通过限制性内切酶酶切、去磷酸化、连接等基因重组技术,将增强型绿色荧光蛋白(Enhanced Green Fluorescent Protein,EGFP)基因和来自莴苣霜霉菌(Bremia lactucae)的启动子(ham34)、终止子重组到真核表达载体pcDNA3.1(-)/hygro中,经大肠杆菌转化后对转化子进行了酶切验证,为大豆疫霉菌遗传转化体系的建立提供载体。
Construction of recombinant vector is the basis for genetic transformation. Recombinant vector used for genetic transformation of Phytophthora sojae was constructed with pcDNA3.1（-）/hygro as the backbone. Enhanced green fluorescent protein （EGFP） gene, the promoter and the terminator of the ham34 gene of Bremia lactucae were recombined to pcDNA3.1（-）/hygro by procedures such as digestion with restriction enzyme, dephosphatasing with CIPA, and lingation with T4 lingase, etc, then transformed into E.coli competent cell. The recombinant plasmid was identified by restriction endonuclease enzyme analysis. It provided a vector for further genetic transformation of Phytophthora sojae.
出处
《东北农业大学学报》
CAS
CSCD
北大核心
2009年第1期9-12,共4页
Journal of Northeast Agricultural University
基金
公益性行业(农业)科研专项(nyhyzx07-053)
黑龙江省"十一五"重点项目(GB06B105-1)
