Reynen M, Reipen I, Sahm H, Sprenger G A
Institut für Biotechnologie 1, Forschungszentrum Jülich GmbH, Federal Republic of Germany.
Mol Gen Genet. 1990 Sep;223(2):335-41. doi: 10.1007/BF00265073.
A set of vectors was constructed for the cloning and expression of heterologous genes in the Gram-negative bacterium Zymomonas mobilis under the control of the pdc promoter of Z. mobilis. The vectors pPTZ1, pPTZ3, and pPTZ4 are based on the cryptic Z. mobilis plasmid pZM02 and on parts of the Escherichia coli plasmids pKK223-3 and pBR322 together with the multiple cloning site of phage M13mp18. DNA fragments can be readily inserted immediately downstream from the pdc promoter at unique restriction sites for KpnI, XbaI and PstI in pPTZ1 and additionally for SmaI and BamHI in pPTZ3. In pPTZ4, the 5' terminal codons of pdc were deleted allowing the formation of gene fusions. Expression of a promoterless chloramphenicol acetyltransferase gene (cat) controlled by the pdc gene promoter resulted in enzyme activities of up to 5.5 U/mg total cell protein in Z. mobilis cells.
构建了一组载体，用于在运动发酵单胞菌的pdc启动子控制下，在革兰氏阴性菌运动发酵单胞菌中克隆和表达异源基因。载体pPTZ1、pPTZ3和pPTZ4基于隐蔽的运动发酵单胞菌质粒pZM02、大肠杆菌质粒pKK223 - 3的部分片段、pBR322以及噬菌体M13mp18的多克隆位点。DNA片段可通过pPTZ1中KpnI、XbaI和PstI的独特限制性位点，以及pPTZ3中额外的SmaI和BamHI，直接插入到pdc启动子下游。在pPTZ4中，删除了pdc的5'末端密码子，从而允许形成基因融合体。由pdc基因启动子控制的无启动子氯霉素乙酰转移酶基因（cat）的表达，在运动发酵单胞菌细胞中产生的酶活性高达5.5 U/mg总细胞蛋白。
