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植物研究 ›› 2006, Vol. 26 ›› Issue (4): 470-474.doi: 10.7525/j.issn.1673-5102.2006.04.001

• 论文 • 上一篇    下一篇

影响农杆菌介导白细胞介素-2基因转化番茄的因素研究

葛艳辉1,2;崔继哲1*;于丽杰1;宋文华2   

  1. (1.哈尔滨师范大学,哈尔滨 150080) (2.天津理工大学,天津 300191)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-07-20 发布日期:2006-07-20
  • 通讯作者: 崔继哲
  • 基金资助:
     

Factors influencing Agrobacterium mediated transformation of interleukin-2 gene to tomato

GE Yan-Hui;CUI Ji-Zhe*;YU Li-Jie;SONG Wen-Hua   

  1. (1.Harbin Normal University, Harbin 150080) (2.Tianjin University of Technology, Tianjin 300191)
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-07-20 Published:2006-07-20
  • Contact: CUI Ji-Zhe
  • Supported by:
     

摘要: 利用农杆菌介导法将白细胞介素-2基因(il-2)导入番茄中,对影响其转化的因素进行了分析。结果表明:农杆菌菌种(EHA105和C58C1)、外植体类型(子叶和下胚轴)、带有不同筛选标记(Kanr、PPTr、Hygr)的载体质粒几个因素对芽诱导分化及转化均有影响。实验共接种转化2018个子叶和下胚轴外植体,获得了47株抗性再生株,对其进行il-2的PCR扩增检测,有44株呈阳性。PCR-Southern杂交证实PCR结果可靠,显示il-2基因已导入到番茄中。

关键词: 白细胞介素-2基因(il-2), 农杆菌介导法, 番茄, 遗传转化

Abstract: An efficient Agrobacterium-mediated transformation system for transferring Interleukin-2 gene (il-2) into tomato was developed by studying several factors that affect the frequency of shoot differentiation and the efficiency of Agrobacterium mediated transformation of explant tissues. Two Agrobacterium strains (EHA105 and C58C1), two types of explant (cotyledon and hypocotyls)and three vectors with different selectable markers (Kanr, PPTr and Hygr) were tested for their influences on transformation. Total 2018 cotyledon and hypocotyl explants from three tomato cultivars were transformed by co-cultivation with Agrobacterium, and 47 selective agent-resistant plants were obtained. 44 out of those were il-2 positive by PCR assay. PCR-Southern hybridization of the PCR-tested plants confirmed the PCR assay, indicating that the il-2 gene had been introduced into tomato.

Key words: interleukin-2 gene, Agrobacterium-mediated method, Lycopersicon esculentum, genetic transformation

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