Blum P, Holzschu D, Kwan H S, Riggs D, Artz S
Department of Microbiology, University of California, Davis 95616.
J Bacteriol. 1989 Jan;171(1):538-46. doi: 10.1128/jb.171.1.538-546.1989.
We have developed an allele exchange system for shuttling sequences of DNA to and from their original chromosomal loci. Cloned segments of the histidine operon of Salmonella typhimurium and the lactose operon of Escherichia coli served as target sequences and were used to develop the system. Replacement and retrieval of target sequences used the phage M13mp vectors and proceeded through an M13 lysogen intermediate. The intermediates and products of allele exchange were characterized by genetic and hybridization analyses. Several unique properties of M13 lysogens were exploited to devise positive selections to detect integration and excision. These positive selections were used to manipulate phenotypically silent alleles.
我们已经开发出一种等位基因交换系统,用于在DNA序列与其原始染色体位点之间来回穿梭。鼠伤寒沙门氏菌组氨酸操纵子和大肠杆菌乳糖操纵子的克隆片段用作靶序列,并用于开发该系统。靶序列的替换和检索使用噬菌体M13mp载体,并通过M13溶原中间体进行。通过遗传分析和杂交分析对等位基因交换的中间体和产物进行了表征。利用M13溶原菌的几个独特特性设计了阳性选择,以检测整合和切除。这些阳性选择被用于操纵表型沉默的等位基因。