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源自原生质体的基因转化玉米植株

Genetically transformed maize plants from protoplasts.

作者信息

Rhodes C A, Pierce D A, Mettler I J, Mascarenhas D, Detmer J J

机构信息

Sandoz Crop Protection Corporation, Palo Alto, CA 94304-1104.

出版信息

Science. 1988 Apr 8;240(4849):204-7. doi: 10.1126/science.2832947.

Abstract

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.

摘要

通过用重组DNA处理原生质体获得了基因转化的玉米植株。从玉米自交系A188的胚性细胞悬浮培养物中消化得到的原生质体,与含有在花椰菜花叶病毒35S启动子区域旁边编码新霉素磷酸转移酶(NPT II)基因的质粒DNA相结合。对原生质体施加高压电脉冲,然后将其在置于玉米悬浮细胞(黑墨西哥甜玉米)饲养层上的滤膜上培养,并在卡那霉素存在下选择生长。筛选出的细胞系显示出NPT II活性。从转化的细胞系中再生出植株并生长至成熟。对从愈伤组织和植株中提取的DNA进行的Southern分析表明存在NPT II基因。

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