Muronets E M, Dubeĭkovskiĭ A N, Kameneva S V
Mol Gen Mikrobiol Virusol. 1987 Feb(2):12-6.
The plasmid pACYC184 is shown to be mobilized for conjugal transfer in Escherichia coli cells by the deleted (Tn7-TcR) derivatives of the hybrid conjugative plasmid pAS8-121 (RP4-Co1E1). Both the mobilized and mobilizing plasmids are autonomously inherited by the recipient cells when the mobilizing plasmid carries single copy of IS8 (the plasmid pAS8-121 delta 16). Cointegrates pAS8-121 delta 16D:: ::pACYC184 are found in the recipient cells with pACYC184 being inserted between two repeats of IS8 if the derivate plasmid pAS8-121 delta 16D having the duplication of IS8 is used to mobilize pACYC184 for conjugal transfer. The insertion of pACYC184 between IS8 repeats in the plasmid pAS8-121 delta 16D eliminates the plasmid ability to be inserted with high frequency into the chromosome of the phototrophic bacterium R. sphaeroides 2R. The cointegrate pAS8-121 delta 16D:: pACYC184 is stable but can be resolved during the transformation deriving the plasmid pACYC184:: IS8. The latter may be used as a probe for isolation and analysis of IS8 DNA sequences and for constructing the vectors on the basis of pACYC184.
已证明质粒pACYC184可被杂种接合质粒pAS8 - 121(RP4 - Co1E1)的缺失(Tn7 - TcR)衍生物在大肠杆菌细胞中进行接合转移。当动员质粒携带单拷贝IS8(质粒pAS8 - 121 delta 16)时,被动员的质粒和动员质粒均可被受体细胞自主遗传。如果使用具有IS8重复序列的衍生质粒pAS8 - 121 delta 16D来动员pACYC184进行接合转移,则在受体细胞中可发现共整合体pAS8 - 121 delta 16D:::pACYC184,其中pACYC184插入在IS8的两个重复序列之间。pACYC184插入质粒pAS8 - 121 delta 16D的IS8重复序列之间会消除该质粒高频插入光合细菌球形红杆菌2R染色体的能力。共整合体pAS8 - 121 delta 16D::pACYC184是稳定的,但在转化过程中可解离产生质粒pACYC184::IS8。后者可作为探针用于分离和分析IS8 DNA序列以及基于pACYC184构建载体。