一种用于快速构建fpg基因的新型三引物PCR方法（TP-PCR）。

A novel tri-primer PCR method (TP-PCR) for rapid construction of fpg gene.

作者信息

Chen Ying Xu, Liu He, Zhang Wen Bo, Jin Yong Feng

机构信息

Department of Environmental Engineering, Zhejiang University, Hangzhou 310029, China.

出版信息

J Microbiol Methods. 2004 Mar;56(3):359-64. doi: 10.1016/j.mimet.2003.11.009.

DOI:10.1016/j.mimet.2003.11.009

PMID:14967227

Abstract

A novel tri-primer polymerase chain reaction method (TP-PCR) was developed for the construction of a fused fpg gene, in which no endonuclease and ligase were used. Instead, two templates and three specifically designed primers were applied. Results showed that pheB and gfp genes, which encodes the catechol 2, 3-dioxygenase and the green fluorescent protein (GFP), respectively, were successfully fused into an fpg gene through the rapid TP-PCR system, indicating that TP-PCR method could be a useful tool for DNA fragment fusion in which no proper endonuclease sites were available.

摘要

开发了一种新型三引物聚合酶链反应方法（TP-PCR）用于构建融合的fpg基因，该方法不使用内切核酸酶和连接酶。相反，应用了两个模板和三个专门设计的引物。结果表明，分别编码儿茶酚2,3-双加氧酶和绿色荧光蛋白（GFP）的pheB和gfp基因通过快速TP-PCR系统成功融合到fpg基因中，这表明TP-PCR方法可能是在没有合适内切核酸酶位点的情况下进行DNA片段融合的有用工具。