Czernilofsky A P, Hain R, Herrera-Estrella L, Lörz H, Goyvaerts E, Baker B J, Schell J
DNA. 1986 Apr;5(2):101-13. doi: 10.1089/dna.1986.5.101.
To compare the effects of different transformation methods on the integration behavior and structural stability of integrated foreign genes in plant cells, tobacco protoplasts were transformed with Escherichia coli plasmid pLGV2103neo DNA using the Ca phosphate DNA coprecipitation technique. Parallel transformations were done by cocultivation with Agrobacterium tumefaciens harboring the Ti plasmid derivatives pGV3850::2103neo or pGV3850::1103neo. A comparison of the fine structure of the integrated donor DNA obtained by direct gene transfer and by cocultivation indicates that the donor DNA in cells transformed by the former technique undergoes structural changes and concatemerizations, while the DNA integrated by the latter procedure is often unaltered. The cotransformed nopaline synthase gene, which is present in the donor Ti plasmid DNA, was inactivated in two out of nine cases. Once integrated, the arrays of selectable marker DNA appear to be structurally stable under different cell culture and selection conditions, as well as after genetic transmission.
为了比较不同转化方法对植物细胞中整合的外源基因的整合行为和结构稳定性的影响，使用磷酸钙DNA共沉淀技术，用大肠杆菌质粒pLGV2103neo DNA转化烟草原生质体。通过与携带Ti质粒衍生物pGV3850::2103neo或pGV3850::1103neo的根癌农杆菌共培养进行平行转化。对通过直接基因转移和共培养获得的整合供体DNA的精细结构的比较表明，通过前一种技术转化的细胞中的供体DNA会发生结构变化和串联，而后一种方法整合的DNA通常未改变。存在于供体Ti质粒DNA中的共转化胭脂碱合成酶基因在9例中有2例失活。一旦整合，可选择标记DNA阵列在不同的细胞培养和选择条件下以及遗传传递后似乎在结构上是稳定的。
