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低能氩离子束介导将绿色荧光蛋白基因导入小麦的研究
引用本文:吴丽芳,李红,宋道军,吴李君,余增亮.低能氩离子束介导将绿色荧光蛋白基因导入小麦的研究[J].南京农业大学学报,2000,23(3):17-20.
作者姓名:吴丽芳  李红  宋道军  吴李君  余增亮
作者单位:中国科学院等离子体物理研究所离子束生物工程实验室,合肥,230031
基金项目:国家自然科学基金!(119890 30 0 ),国家重点科技攻关项目!(96 5 38 0 2 0 1)
摘    要:用低能氩离子束介导将改良的绿色荧光蛋白基因(mGFP4)导入小麦栽培品种皖9210和皖麦32号的成熟胚细胞。在含有100 ̄140m/L巴龙霉素培养基上继代培养后,获得了一批抗性愈伤组织。经过分化培养,皖9210获得5株再生苗,皖麦32号获得32株绿苗,对照的200枚成熟胚未获得再生苗。对再生苗进行PCR检测,均扩增出600bp的nptⅡ基因片段。取4株长势好的绿苗进行Southern杂交分析,结果

关 键 词:离子束  成熟胚  绿色荧光蛋白基因

Study on transferring improved green fluorescent protein gene into wheat via low energy Ar+ implantation
Wu Lifang,Li Hong,Song Daojun,Wu Lijun,Yu Zengliang.Study on transferring improved green fluorescent protein gene into wheat via low energy Ar+ implantation[J].Journal of Nanjing Agricultural University,2000,23(3):17-20.
Authors:Wu Lifang  Li Hong  Song Daojun  Wu Lijun  Yu Zengliang
Abstract:An improved GFP gene(mGFP4)was introduced into mature embryo cells of wheat cultivars Wan 9210 and Wanmai 32 via low energy ion beam mediated delivery technique.Resistant calli were selected on medium containing paromomycin(100?140 mg/L).Five green plants were regenerated from resistant calli of Wan 9210 derived from 387 implanted mature embryos.32 green plants were obtained from 776 irradiated mature embryos in Wanmai 32.No green plant was regenerated from calli of 200 non transformed embryos.PCR assays of 37 green plants showed that they all obtained the expected size of amplified DNA fragment(600 bp).Southern blot of 4 well?developed green plants confirmed stable integration of GFP gene into wheat genome.The average transformation frequencies of Wan 9210 and Wanmai 32 were 1 3% and 4 1%,respectively,according to the results of PCR assays.
Keywords:ion beam  mature embryos  green fluorescent protein gene  transgenic wheat plants
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