Kadokawa Y, Kusakabe T, Kamachi Y, Isobe K, Kondoh H, Ohyama T
Division of Molecular Biology, Meiji Institute of Health Science, Odawara, Japan.
Gene. 1995 Feb 14;153(2):277-8. doi: 10.1016/0378-1119(94)00803-z.
A new vector, pATO, was constructed for rapid cloning and analysis of eukaryotic promoters. When a recombinant pATO, carrying a promoter sequence in its multiple cloning site, was introduced into COS cells, Thy-1.2 protein was produced on the cell surface, and was easily identified by an fluorescein-conjugated anti-Thy-1.2 antibody. The intensity of the fluorescence reflected the strength of the inserted promoter. Since pATO could replicate efficiently in COS cells, the recombinant plasmids recovered from a single COS cell were sufficient to transform Escherichia coli cells. This plasmid is applicable for the rapid and labor saving cloning of promoter elements.
构建了一种新载体pATO，用于真核启动子的快速克隆和分析。当一个在其多克隆位点携带启动子序列的重组pATO被导入COS细胞时，细胞表面会产生Thy-1.2蛋白，并且可以通过荧光素偶联的抗Thy-1.2抗体轻松识别。荧光强度反映了插入启动子的强度。由于pATO能够在COS细胞中高效复制，从单个COS细胞回收的重组质粒足以转化大肠杆菌细胞。该质粒适用于启动子元件的快速且省力的克隆。
