一种用于鉴定18种转基因大豆转化体的多靶标质粒的构建与应用

被引:7
作者
史宗勇 [1 ]
陈子言 [2 ]
祁琛 [1 ]
王成 [1 ]
赵梦晓 [1 ]
李夏莹 [2 ]
王文斌 [1 ]
袁建琴 [1 ]
许冬梅 [1 ]
乔永刚 [1 ]
刘建东 [1 ]
张秀杰 [2 ]
高建华 [1 ]
机构
[1] 山西农业大学生命科学学院
[2] 农业农村部科技发展中心
关键词
转基因大豆; 质粒; 阳性物质; 转化体特异性检测; 转化体特征序列;
D O I
暂无
中图分类号
S565.1 [大豆];
学科分类号
摘要
本文拟构建一种可满足我国转基因大豆转化体鉴定需求的多靶标质粒(multiple target plasmid, MTP)分子。根据相应的国家农业检测标准,选定我国已经批准进口的14种独立转化体和尚未批准但具有重要应用前景的4种重要转化体,将这18种转化体的特征序列以及大豆内标准基因Lectin的检测序列合理排列后,接入pUC18质粒,构建一种多靶标质粒pDDID-1905。在插入序列之间,分布着多种位点单一的限制性内切酶识别序列,可用于后期对该质粒的修订和更新。对pDDID-1905质粒中包含的19种靶标序列进行聚合酶链式反应检测,以验证该质粒的实用性。结果显示,所有靶标序列均被成功扩增,其大小与预期一致。总之,本文构建的适用于18种转基因大豆转化体鉴定的阳性质粒分子pDDID-1905,是目前包含大豆转化体特征序列最多的多靶标质粒,其构建和应用将极大地简化烦琐的阳性物质准备工作。
引用
收藏
页码:280 / 290
页数:11
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