Takahashi T, Ryan K W, Portner A
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101-0318.
Genet Anal Tech Appl. 1992 Jun;9(3):91-5. doi: 10.1016/1050-3862(92)90004-o.
To facilitate the construction of recombinant plasmids for expressing cloned genes with T7 RNA polymerase supplied by recombinant vaccinia virus, a plasmid expression vector was designed by combining parts of plasmids pTZ18R, pBluescript II KS+, and pAR2529. The 3043-bp plasmid pTF1 has a T7 RNA polymerase promoter, multiple cloning site for insertion of foreign genes, and a T7-specific transcription termination signal. Plasmid pTF1 had several advantages compared with the reference plasmid pAR2529, including more efficient replication in bacteria, greater flexibility in the insertion and subcloning of foreign genes, and increased efficiency of liposome-mediated introduction into cultured cells for expression of the foreign gene.
为便于构建用于表达由重组痘苗病毒提供的T7 RNA聚合酶克隆基因的重组质粒，通过组合质粒pTZ18R、pBluescript II KS+和pAR2529的部分片段设计了一种质粒表达载体。3043 bp的质粒pTF1具有T7 RNA聚合酶启动子、用于插入外源基因的多克隆位点以及T7特异性转录终止信号。与参考质粒pAR2529相比，质粒pTF1具有多个优点，包括在细菌中更高效的复制、在外源基因插入和亚克隆方面更大的灵活性，以及提高脂质体介导导入培养细胞中外源基因表达的效率。
