Tangney M, Jørgensen P L, Diderichsen B, Jørgensen S T
Department of Bacterial Gene Technology, Novo Nordisk A/S, Copenhagen, Denmark.
FEMS Microbiol Lett. 1995 Jan 1;125(1):107-14. doi: 10.1111/j.1574-6968.1995.tb07343.x.
We have developed a strategy for the integration and stable amplification of DNA sequences in the chromosome of poorly transformable bacilli, which avoids the presence of a functional plasmid replication system in the integrated DNA. The parental vector for integration contains two plus origins of replication from pUB110 in the same orientation on a single plasmid. Due to the direct repeats, such plasmids produce two individual progeny vectors, one of which is dependent on the other for replication, as it lacks a functional rep gene. We have used such a progeny vector system to integrate and amplify DNA on the chromosome of Bacillus licheniformis, and show that the structure is stable in the absence of selective pressure.
我们已经开发出一种策略，用于在难以转化的杆菌染色体中整合和稳定扩增DNA序列，该策略避免了整合DNA中存在功能性质粒复制系统。用于整合的亲本载体在单个质粒上以相同方向包含来自pUB110的两个正向复制起点。由于存在同向重复序列，此类质粒会产生两个单独的子代载体，其中一个子代载体因缺乏功能性rep基因，其复制依赖于另一个子代载体。我们已使用这样的子代载体系统在地衣芽孢杆菌染色体上整合和扩增DNA，并表明在无选择压力的情况下该结构是稳定的。
