%0 Journal Article
%T 运用Gibson克隆技术构建蒺藜苜蓿成熟根瘤的酵母AD-cDNA文库<br>Applying Gibson cloning technique in constructing cDNA library
%A 陈琳
%A 余海翔
%A 张忠明
%J 华中农业大学学报
%D 2018
%X 以蒺藜苜蓿（A17）的成熟根瘤为材料，联合应用Gibson克隆技术和SMART cDNA技术构建cDNA文库，其转化效率达到7.74×105 cfu/3 μg pGADT7-Rec-New。随机菌落PCR检测结果表明，插入DNA片段的大小为0.5 ~2.0 kb，其中插入片段＞0.75 kb的占45.6%，文库的重组效率达到91.7%，构建的cDNA文库以质粒形式保藏备用。采用此法不仅可以构建用于酵母单杂交或双杂交的文库，同样也适用于其他质粒文库的构建。利用酵母单杂交技术，以在根瘤中高表达基因启动子元件为诱饵，筛选构建的文库，得到拟似与DNA互作的蛋白质。<br>The mature nodules of Medicago truncatula (A17) were used to construct a cDNA library with Gibson cloning technique and SMART cDNA library construction technique.The result of library testing showed that the transformation efficiency of the library was 7.74×105 transformants/3 μg pGADT7-Rec-New.Some colonies were randomly picked to run PCR.Results showed that the inserted DNA fragment size was ranged from 0.5 kb to 2.0 kb,with the recombinant rate of 91.7%.The cDNA library was stored as plasmid.This method can be used to construct library for yeast one or double hybrid and other plasmid library.The library was used to find protein(TF) that can interact with the DNA fragment of interest and some TFs were obtained.
%K 蒺藜苜蓿 根瘤 AD-cDNA文库 Gibson克隆技术&lt
%K br&gt
%K Medicago truncatula nodule AD-cDNA library Gibson cloning technique
%U http://hnxbl.cnjournals.net/hznydxzr/ch/reader/view_abstract.aspx?file_no=20180204&flag=1