Stepanov A I, Urlapova S V, Zimina M S
Genetika. 1979 Nov;15(11):1971-8.
Hybrid plasmid pAS8, that consists of RP4 and ColE1 replicons, is incompatible with RP4 but co-exists with ColE1 replicon. The deletion mutants of pAS8, that replicates under the control of ColE1 replicon only, are incompatible with ColE1 derivatives, although the copy number of pAS8 and its deletion mutants in the cell is the same (1-3 per the chromosome). Incompatibility effect of plasmids is expressed in a sharp decrease in transformant's yield during selection of incoming and resident plasmids markers. In this case incompatibility is a very fast process, that leads to the elimination of resident or incoming plasmid just before plating on the selective medium. On the base of negative control theory, the repressors yield, synthesized in the presence of 1-3 copies of the deletion mutant is enough for the expression of ColE1-specific incompatibility. This ColE1 incompatibility is probably connected with the functional activity of ColE1 replicon. When ColE1 factor replicates under the control of RP4 replicon the expression of ColE1-specific incompatibility does not occur. Possibly, the incompatibility effect takes place when pAS8 deletion mutants cause the synthesis of ColE1-specific repressor. Also, the replicons of ColE1 may competein the membrane attachment site.
由RP4和ColE1复制子组成的杂种质粒pAS8与RP4不相容,但与ColE1复制子共存。仅在ColE1复制子控制下复制的pAS8缺失突变体与ColE1衍生物不相容,尽管pAS8及其缺失突变体在细胞中的拷贝数相同(每条染色体1 - 3个)。在选择导入质粒和常驻质粒标记时,转化体产量急剧下降体现了质粒的不相容效应。在这种情况下,不相容是一个非常快速的过程,导致在铺板到选择培养基之前常驻质粒或导入质粒被消除。基于负调控理论,在存在1 - 3个拷贝的缺失突变体时合成的阻遏物产量足以表达ColE1特异性不相容性。这种ColE1不相容性可能与ColE1复制子的功能活性有关。当ColE1因子在RP4复制子控制下复制时,不会发生ColE1特异性不相容性的表达。可能当pAS8缺失突变体导致ColE1特异性阻遏物合成时,不相容效应就会发生。此外,ColE1的复制子可能在膜附着位点存在竞争。