Williams J G, Szalay A A
Gene. 1983 Sep;24(1):37-51. doi: 10.1016/0378-1119(83)90129-4.
The blue-green alga, Synechococcus R2, is transformed to antibiotic resistance by chimeric DNA molecules consisting of Synechococcus R2 chromosomal DNA linked to antibiotic-resistance genes from Escherichia coli. Chimeric DNA integrates into the Synechococcus R2 chromosome by homologous recombination. The efficiency of transformation, as well as the stability of integrated foreign DNA, depends on the position of the foreign genes relative to Synechococcus R2 DNA in the chimeric molecule. When the Synechococcus R2 DNA fragment is interrupted by foreign DNA, integration occurs through replacement of chromosomal DNA by homologous chimeric DNA containing the foreign insert; transformation is efficient and the foreign gene is stable. Mutagenesis in some cases attends integration, depending on the site of insertion. Foreign DNA linked to the ends of Synechococcus R2 DNA in a circular molecule, however, integrates less efficiently. Integration results in duplicate copies of Synechococcus R2 DNA flanking the foreign gene and the foreign DNA is unstable. Transformation in Synechococcus R2 can be exploited to modify precisely and extensively the genome of this photosynthetic microorganism.
蓝绿藻聚球藻R2通过由聚球藻R2染色体DNA与来自大肠杆菌的抗生素抗性基因相连组成的嵌合DNA分子转化为抗生素抗性。嵌合DNA通过同源重组整合到聚球藻R2染色体中。转化效率以及整合的外源DNA的稳定性取决于嵌合分子中外源基因相对于聚球藻R2 DNA的位置。当聚球藻R2 DNA片段被外源DNA中断时,整合通过含有外源插入片段的同源嵌合DNA取代染色体DNA而发生;转化效率高且外源基因稳定。在某些情况下,诱变伴随着整合,这取决于插入位点。然而,在环状分子中与聚球藻R2 DNA末端相连的外源DNA整合效率较低。整合导致外源基因两侧出现聚球藻R2 DNA的重复拷贝,并且外源DNA不稳定。聚球藻R2中的转化可用于精确而广泛地修饰这种光合微生物的基因组。