[绿色荧光标记重组质粒的构建及其在粉红粘帚霉中的应用]
[Construction of a recombination plasmid labelled with green fluorescence and its application in Clonostachys rosea].
作者信息
Niu Qiuhong, Ke Tao, Zhang Lin, Hui Fengli, Chen Jibao
机构信息
School of Life Science and Technology, Nanyang Normal University, Nanyang 473061, China.
出版信息
Wei Sheng Wu Xue Bao. 2011 Jul;51(7):991-7.
OBJECTIVE
We constructed a recombinant eukaryotic expression vector harboring green fluorescent protein (GFP) and the hygromycin resistance gene hph, and observed its expression in Clonostachys rosea.
METHODS
We used PCR, enzyme digestion, phosphorylation, ligation and transformation to construct the plasmid. Using protoplast preparation and transformation technologies, we expressed the plasmid in the fungi C. rosea.
RESULTS
We created the eukaryotic expression vector, transformed it into C. rosea and observed green fluorescence with fluorescence microscope.
CONCLUSION
The successful construction of the pANGH3 recombinant plasmid and its expression in C. rosea establishes a new model for studying fungal infection mechanisms.
目的
构建携带绿色荧光蛋白（GFP）和潮霉素抗性基因hph的重组真核表达载体，并观察其在粉红粘帚霉中的表达情况。
方法
采用聚合酶链反应（PCR）、酶切、磷酸化、连接和转化等方法构建质粒。利用原生质体制备和转化技术，在粉红粘帚霉菌中表达该质粒。
结果
成功构建了真核表达载体，并将其转化到粉红粘帚霉中，通过荧光显微镜观察到绿色荧光。
结论
pANGH3重组质粒的成功构建及其在粉红粘帚霉中的表达，为研究真菌感染机制建立了新的模型。
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