Friedberg D, Seijffers J
Mol Gen Genet. 1986 Jun;203(3):505-10. doi: 10.1007/BF00422077.
This study presents plasmid systems that utilize regulatory signals of bacteriophage Lambda to accomplish regulated expression of cloned genes in an A. nidulans R2 derivative strain. An operator-promoter region and the temperature-sensitive repressor gene cI857 of bacteriophage Lambda were employed. Linked to a cyanobacterial replicon, the plasmid vectors efficiently transformed Anacystis and were stably maintained within this host. The cat structural gene, encoding chloramphenicol acetyltransferase, was used to demonstrate that expression can be regulated by temperature shift. We have identified in extracts from the vector bearing Anacystis, a protein similar in size and immunology to the Lambda repressor. The systems described should allow controlled expression of adventitious genes in the cyanobacterial host.
本研究提出了利用噬菌体λ调控信号在构巢曲霉R2衍生菌株中实现克隆基因调控表达的质粒系统。采用了噬菌体λ的一个操纵子 - 启动子区域和温度敏感型阻遏基因cI857。与蓝细菌复制子相连,这些质粒载体有效地转化了集胞藻,并在该宿主内稳定维持。编码氯霉素乙酰转移酶的cat结构基因被用于证明表达可通过温度变化进行调控。我们在携带集胞藻的载体提取物中鉴定出一种在大小和免疫学性质上与λ阻遏蛋白相似的蛋白质。所描述的系统应能在蓝细菌宿主中实现外源基因的可控表达。