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地高辛标记LEA3蛋白基因探针的制备及其在转基因草莓核酸杂交中的应用

Preparation of LEA3 Protein Gene Probe Labeled by Digoxigenin and Application in Detection of Transgenic Strawberry

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【作者】 王俊丽葛会波彭士琪张红梅续九如

【Author】 Wang Junli, Ge Huibo, Peng Shiqi, Zhang Hongmei,and Xu Jiuru(College of Life Sciences , Hebei University ,Baoding 071002 China; College of Biological Sciences and Technology, Beijing Forestry U niversity, Beijing, 100083 China); College of Horticulture , Hebei Agricultural University, Baoding, Hebei 071000 China)

【机构】 河北大学生命科学学院河北农业大学园艺学院北京林业大学生物科学与技术学院 河北 保定 071002 北京林业大学生物科学与技术学院北京 100083河北 保定 071000河北 保定 071002北京 100083

【摘要】 用地高辛对胚胎发生晚期丰富蛋白LEA3基因片段(1.0 kh)进行了探针标记。斑点杂交表明,该探针的检测灵敏度为5 Pg。用地高辛标记和未标记的标准分子量DL 2000进行杂交时,只检测到2.00、1.00、0.75、0.50 kb4条杂交带。应用LEA3探针对转基因草莓(Fragaria ananassa Duch.)进行了Southern blot分析,检测到1.0 kh杂交带,表明外源LEA3基因整合到草莓染色体上。

【Abstract】 Plasmid DNA probe of late embryogenesis abundant protein gene, LEA3 (1.0 kb), was labeled by digoxigenin. Dot blot showed that the probe was sensitive, able to detect 5 pg homologous DNA. Four hybridization bands (2.0 kb, 1.0 kb, 0.75 kb and 0.5 kb) were detected in Southern blot analysis of marker DL2000. The LEA3 probe labeled by digoxigenin was used in Southern blot analysis of transgenic strawberry (Fragaria ananassa Duch.) and hybridization bands were detected. The result indicated that LEA3 gene was integrated into the genome of strawberry.

【基金】 河北省自然科学基金资助项目(302110)。
  • 【文献出处】 果树学报 ,Journal of Fruit Science , 编辑部邮箱 ,2004年04期
  • 【分类号】S668.4
  • 【被引频次】4
  • 【下载频次】219
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