GENETICALLY TRANSFORMED MAIZE PLANTS FROM PROTOPLASTS
GENETICALLY TRANSFORMED MAIZE PLANTS FROM PROTOPLASTS
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DOI：
10.1126/science.2832947
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发表时间：
1988-04-08
期刊：
影响因子：
56.9
通讯作者：
DETMER, JJ
中科院分区：
文献类型：
--
作者：
RHODES, CA;PIERCE, DA;DETMER, JJ
用重组DNA处理原生质体获得转基因玉米植株。从玉米自交系A188胚性细胞悬浮培养中消化的原生质体与含有新霉素磷酸转移酶（NPT II）基因的质粒DNA在花菜花叶病毒35S启动子区附近结合。对原生质体施加高压电脉冲，然后在放置在玉米悬浮细胞（黑墨西哥甜）饲养层上的过滤器上生长，并选择在卡那霉素存在下生长。所选细胞系显示NPTⅱ活性。植物从转化的细胞系中再生并生长到成熟。从愈伤组织和植株中提取的DNA的Southern分析表明存在NPT II基因。
Genetically transformed maize plants were obtained from protoplasts treated with recombinant DNA. Protoplasts that were digested from embryogenic cell suspension cultures of maize inbred A188 were combined with plasmid DNA containing a gene coding for neomycin phosphotransferase (NPT II) next to the 35S promoter region of cauliflower mosaic virus. A high voltage electrical pulse was applied to the protoplasts which were then grown on filters placed over feeder layers of maize suspension cells (Black Mexican Sweet) and selected for growth in the presence of kanamycin. Selected cell lines showed NPT II activity. Plants were regenerated from transformed cell lines and grown to maturity. Southern analysis of DNA extracted from callus and plants indicated the presence of the NPT II gene.