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通过体细胞胚胎发生再生系统经微粒轰击获得的转基因花生植株。

Transgenic peanut plants obtained by particle bombardment via somatic embryogenesis regeneration system.

作者信息

Deng X Y, Wei Z M, An H L

机构信息

Institute of Plant Physiology and Ecology, Chinese Academy Sciences, Shangha.

出版信息

Cell Res. 2001 Jun;11(2):156-60. doi: 10.1038/sj.cr.7290081.

Abstract

After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resistance to hygromycin and a chimeric intron-gus gene. Selection for hygromycin resistant calluses and somatic embryos was initiated at 10th d post-bombardment on medium containing 10-25 mg/L hygromycin. Under continuous selection, hygromycin resistant plantlets were regenerated from somatic embryos and were recovered from nearly 1.6% of the bombarded cotyledons. The presence and integration of foreign DNA in regenerated hygromycin resistant plants was confirmed by PCR (polymerase chain reaction) for the intron-gus gene and by Southern hybridization of the hph gene. GUS enzyme activity was detected in leaflets from transgenic plants but not from control, non-transformed plants. The production of transgenic plants are mainly based on a newly improved somatic embryogenesis regeneration system developed by us.

摘要

经过预培养和渗透处理后,利用含有赋予潮霉素抗性的嵌合hph基因和嵌合内含子 - gus基因的质粒,通过粒子轰击法对未成熟花生(Arachis hypogaea L.)合子胚的子叶进行转化。在轰击后第10天,在含有10 - 25 mg/L潮霉素的培养基上开始筛选抗潮霉素的愈伤组织和体细胞胚。在持续筛选下,从体细胞胚再生出抗潮霉素的植株,并且从近1.6%的轰击子叶中获得了再生植株。通过针对内含子 - gus基因的PCR(聚合酶链反应)和hph基因的Southern杂交,证实了再生的抗潮霉素植株中存在并整合了外源DNA。在转基因植物的小叶中检测到了GUS酶活性,而在对照的未转化植物中未检测到。转基因植物的产生主要基于我们新改进的体细胞胚胎发生再生系统。

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