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用植株转化法将GUS基因导入桑树幼苗的研究
引用本文:陆小平,楼程富,王波,小岛峯雄.用植株转化法将GUS基因导入桑树幼苗的研究[J].蚕业科学,2004,30(2):129-132.
作者姓名:陆小平  楼程富  王波  小岛峯雄
作者单位:浙江大学动物科学学院蚕蜂科学系,杭州,310029;苏州大学农业科学与技术学院;浙江大学动物科学学院蚕蜂科学系,杭州,310029;苏州大学农业科学与技术学院;日本信州大学纤维学部
摘    要:为探讨简便易行的农杆菌介导的转化方法 ,用工程农杆菌A 110株 (LBA4 4 0 4 /pIG12 1 Hm)对桑幼苗子叶腋隙进行刺伤感染 ,感染部位长出的桑芽表型异常。取其基因组DNA进行分子生物学鉴定 ,结果显示 :以基因组DNA为模板 ,经PCR反应后得到特异性扩增片段用SalⅠ酶解 ,酶切产物与预计大小完全相符 ;用GUS基因作探针进行Southern杂交和用Km基因作探针对基因组DNA进行Southern杂交 ,均获得较强的杂交信号。

关 键 词:  农杆菌介导  整体转化
文章编号:0257-4799(2004)02-0129-04
修稿时间:2003年12月18日

Transducting GUS Gene Into Mulberry Seedling by Plant Transformation Technique
LU Xiaoping , LOU Chenfu WANG Bo MINEO Kojima.Transducting GUS Gene Into Mulberry Seedling by Plant Transformation Technique[J].Acta Sericologica Sinica,2004,30(2):129-132.
Authors:LU Xiaoping  LOU Chenfu WANG Bo MINEO Kojima
Institution:LU Xiaoping 1,2 LOU Chenfu 1 WANG Bo 2 MINEO Kojima 3
Abstract:In order to get convenient manipulative translation technique with Agrobacterium tumefaciens, the cotyledon leaf gap of mulberry seedlings was infected with Agrobacterium tumefaciens A-110 strain (LBA4404/pIG121-Hm) by perforation. It was observed that the mulberry buds on the infected position were abnormal, its genomic DNA was identified by molecular biology method. The results indicated as: The expected fragment was amplified by PCR using the genomic DNA as template, which was digested by Sal Ⅰ, and the fragments were similar to that we expected in size. The PCR product was hybridized with the genomic DNA by Southern bloting using the GUS gene or Km gene as probe, a strong signal was detected.
Keywords:Morus albaTransformation of Agrobacterium tumefaciensIn plant transformation
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