Matsushima P, Broughton M C, Turner J R, Baltz R H
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285.
Gene. 1994 Aug 19;146(1):39-45. doi: 10.1016/0378-1119(94)90831-1.
Cosmid pOJ436, containing large inserts of Saccharopolyspora spinosa (Ss) DNA, was transferred by conjugation from Escherichia coli to Ss an integrated into the chromosome, apparently by homologous recombination, at high frequencies (10(-5) to 10(-4) per recipient). Transfer was mediated by the plasmid RP4 (RK2) transfer functions in E. coli, and the RK2 oriT function located on pOJ436 [Bierman et al., Gene 116 (1992) 43-49]. pOJ436 lacking Ss DNA, or containing a small insert (approx. 2 kb) of Ss DNA, conjugated from E. coli and integrated at either of two bacteriophage phi C31 attB sites at low frequency (approx. 10(-7) per recipient). Exconjugants containing homologous inserts or inserts at the phi C31 attB sites were stable in the absence of antibiotic selection, and most produced control levels of tetracyclic macrolide A83543 factors. Some exconjugants contained similar kinds of large deletions and were defective in macrolide production.
粘粒pOJ436含有多刺糖多孢菌（Ss）DNA的大片段，通过接合作用从大肠杆菌转移至Ss，并显然通过同源重组以高频率（每个受体10^(-5)至10^(-4)）整合到染色体中。转移由大肠杆菌中的质粒RP4（RK2）转移功能介导，且RK2 oriT功能位于pOJ436上[Bierman等人，《基因》116（1992年）43 - 49]。缺乏Ss DNA或含有小片段（约2 kb）Ss DNA的pOJ436从大肠杆菌接合转移，并以低频率（每个受体约10^(-7)）整合到两个噬菌体phi C31 attB位点中的任一个。含有同源插入片段或phi C31 attB位点插入片段的接合子在无抗生素选择的情况下是稳定的，且大多数产生四环大环内酯A83543因子的对照水平。一些接合子含有类似类型的大片段缺失，并且在大环内酯产生方面存在缺陷。
