Deng Z X, Zhou X F
Laboratory of Agricultural Antibiotics, Huazhong Agricultural University, Wuhan.
Chin J Biotechnol. 1990;6(3):165-72.
The 13 kb DNA segment responsible for replication has been isolated from Streptomyces hygroscopicus subsp. yingchengensis strain 10-22 using a pBR322-derived Streptomyces replication-probe vector pIJ2703. The restriction map of the hybrid plasmid, pHZ54, has been constructed. Following a series of deletion and subcloning experiments, the minimal functional region for replication was located within a 2.86 kb DNA fragment. Subsequently, this region was further shortened into a 2 kb region by exonuclease III deletion. A few small bifunctional plasmids able to replicate in E. coli as well as in Streptomyces, were obtained during the above process. Some plasmids could be used as shuttle vectors.
利用源自pBR322的链霉菌复制探针载体pIJ2703，从吸水链霉菌应城变种菌株10 - 22中分离出负责复制的13 kb DNA片段。构建了杂交质粒pHZ54的限制性图谱。经过一系列缺失和亚克隆实验，确定复制的最小功能区域位于一个2.86 kb的DNA片段内。随后，通过核酸外切酶III缺失将该区域进一步缩短至2 kb区域。在上述过程中获得了一些能够在大肠杆菌和链霉菌中复制的小型双功能质粒。一些质粒可作为穿梭载体。
