Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan, ROC.
Biotechnol Prog. 2009 Nov-Dec;25(6):1582-6. doi: 10.1002/btpr.274.
To quickly find an optimal expression system for recombinant protein production, a set of vectors with the same restriction sites were constructed for parallel cloning of a target gene and recombinant protein production in prokaryotic and eukaryotic expression systems, simultaneously. These vectors include nucleotide sequences encoding protein tags and protease recognition sites for tag removal, followed by the cloning sites 5'-EcoRI/3'-XhoI identical in these vectors for ligating with the sticky-end PCR product of a target gene. Our vectors allow parallel gene cloning and protein production in multiple expression systems with minimal cloning effort.
为了快速找到重组蛋白生产的最佳表达系统，我们构建了一组具有相同限制位点的载体，用于同时在原核和真核表达系统中平行克隆目的基因和重组蛋白。这些载体包括编码蛋白标签和蛋白酶识别位点的核苷酸序列，用于标签去除，随后是克隆位点 5'-EcoRI/3'-XhoI，这些载体中的克隆位点相同，用于与目的基因的粘性末端 PCR 产物连接。我们的载体允许在多个表达系统中进行平行基因克隆和蛋白生产，只需最小的克隆工作量。
