Hoffmann T, Golz C, Schieder O
Institute of Applied Genetics, Free University Berlin, Germany.
Curr Genet. 1994 Dec;27(1):70-6. doi: 10.1007/BF00326581.
Different transgenic plants of Brassica napus, Brassica nigra, Datura innoxia and Vicia narbonensis expressing the hph gene under the control of the 35s promoter were co-cultivated with mycelial material of Aspergillus niger in microcosms under sterile conditions. A significantly higher number of hygromycin B-resistant colonies of re-isolated fungi was obtained if compared with co-cultures with non-transgenic plants. The hph gene and other foreign sequences could be detected in some of the resistant strains only for a short time after selection, indicating a rapid loss of foreign DNA. A more stable transgenic strain was obtained after co-culture with transgenic plants of D. innoxia including a high number of hph copies in their genome. DNA with detected pUC sequences was prepared to transform E. coli DH5 alpha. One of the recovered plasmids is shown to include pieces of the plant-transforming vector and a foreign sequence. The 35s-regulated expression of genes is studied in A. niger.
在无菌条件下,将在35s启动子控制下表达潮霉素磷酸转移酶(hph)基因的不同转基因甘蓝型油菜、黑芥、无刺曼陀罗和窄叶野豌豆植株与黑曲霉的菌丝体材料在微观环境中共同培养。与非转基因植物的共培养相比,重新分离的真菌中潮霉素B抗性菌落的数量显著更高。在选择后仅短时间内,在一些抗性菌株中可检测到hph基因和其他外源序列,表明外源DNA迅速丢失。与基因组中包含大量hph拷贝的无刺曼陀罗转基因植株共培养后,获得了一个更稳定的转基因菌株。制备了检测到pUC序列的DNA以转化大肠杆菌DH5α。回收的质粒之一显示包含植物转化载体片段和一个外源序列。在黑曲霉中研究了基因的35s调控表达。