Metapontum Agrobios, I-75010, Metaponto, Italy.
Theor Appl Genet. 1990 Sep;80(3):395-401. doi: 10.1007/BF00210079.
Efficient delivery of genomic DNA fragments to maize protoplasts was obtained by new methods using the polycation Polybrene or Lipofectin cationic liposomes. Stable kanamycin-resistant secondary transformants were recovered after transfection with genomic DNA from a maize cell line that had previously been tagged with the bacterial gene neomycin phosphotransferase (nptII) in a first-round transformation. The frequency of secondary transformants with nptII-homologous DNA sequences was 3% or 6% of all randomly picked microcalli after Polybrene or Lipofectin-mediated transfection, respectively. Transformation with genomic DNA by these methods may allow easy transfer of uncloned genes encoding desirable characteristics to crop species that can be regenerated from protoplasts.
通过使用聚阳离子多聚体或脂质体 Lipofectin 等新方法，实现了将基因组 DNA 片段高效递送至玉米原生质体。通过转染第一轮转化中用细菌基因新霉素磷酸转移酶（nptII）标记的玉米细胞系的基因组 DNA，获得了稳定的卡那霉素抗性二级转化体。在 Polybrene 或 Lipofectin 介导的转染后，随机挑选的微愈伤组织中，分别有 3%或 6%的二级转化体具有 nptII 同源 DNA 序列。通过这些方法用基因组 DNA 进行转化可能会使未克隆的基因（其编码的特性是人们所期望的）更容易转移到可由原生质体再生的农作物物种中。
