Averhoff B, Gregg-Jolly L, Elsemore D, Ornston L N
Department of Biology, Yale University, New Haven, Connecticut 06511.
J Bacteriol. 1992 Jan;174(1):200-4. doi: 10.1128/jb.174.1.200-204.1992.
DNA within Escherichia coli colonies carrying cloned Acinetobacter calcoaceticus genes transforms mutant A. calocaceticus cells with high efficiency. Therefore, E. coli colonies containing such cloned genes can be identified by replica plating onto a lawn of A. calcoaceticus mutant cells. Transformation of A. calcoaceticus also facilitates gap repair and thus allows recovery of specified chromosomal segments in recombinant plasmids. These procedures were used to demonstrate the clustering of A. calcoaceticus genes required for utilization of p-hydroxybenzoate. Chromosomal linkage of the bacterial genes, contained in different operons separated by about 10 kbp of DNA, may have been selected on the basis of their physiological interdependence.
携带克隆的醋酸钙不动杆菌基因的大肠杆菌菌落中的DNA能高效转化突变型醋酸钙不动杆菌细胞。因此,通过将含有此类克隆基因的大肠杆菌菌落影印接种到醋酸钙不动杆菌突变细胞的菌苔上,可以对其进行鉴定。醋酸钙不动杆菌的转化还促进了缺口修复,从而使得重组质粒中特定染色体片段得以回收。这些方法被用于证明利用对羟基苯甲酸所需的醋酸钙不动杆菌基因的聚类。细菌基因位于由约10kbp的DNA分隔的不同操纵子中,其染色体连锁可能是基于它们的生理相互依赖性而被选择的。