GENOMIC DNA CAN BE USED WITH CATIONIC METHODS FOR HIGHLY EFFICIENT TRANSFORMATION OF MAIZE PROTOPLASTS
GENOMIC DNA CAN BE USED WITH CATIONIC METHODS FOR HIGHLY EFFICIENT TRANSFORMATION OF MAIZE PROTOPLASTS
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DOI：
10.1007/bf00210079
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发表时间：
1990-01-01
影响因子：
5.4
通讯作者：
STADLER, J
中科院分区：
文献类型：
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作者：
ANTONELLI, NM;STADLER, J
利用多阳离子聚烯脂质体或脂质体脂质体，实现了玉米原生质体基因组DNA片段的高效传递。在第一轮转化中，先前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转染后，获得了稳定的卡那霉素耐药二级转化子。在Polybrene介导或lipofectin介导转染后，nptii同源DNA序列的次级转化子分别占所有随机挑选的微愈伤组织的3%和6%。用这些方法转化基因组DNA，可以很容易地将编码理想特性的非克隆基因转移到可以从原生质体再生的作物物种中。
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 tranfer of uncloned genes encoding desirable characteristics to crop species that can be regenerated from protoplasts.