转基因油菜Ms8质粒分子的制备

doi:10.3969/j.issn.1004-1524.2017.05.02

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (5): 694-700.DOI: 10.3969/j.issn.1004-1524.2017.05.02

转基因油菜Ms8质粒分子的制备

余笑波¹, 沙跃兵¹, 张江东¹, 赵雷¹, 陈怡¹, 隋志伟²

1.浙江省计量科学研究院,浙江杭州 310018；
2.中国计量科学研究院,北京 100013

收稿日期:2016-12-29 出版日期:2017-05-20 发布日期:2017-06-06
作者简介:余笑波(1985-),男,浙江杭州人,硕士,工程师,主要从事生物化学分析仪器的检定和生物化学相关标准物质研制。E-mail: yuxiaobo.123@163.com
基金资助:
浙江省质量技术监督局一般项目(20150211)

Development of a reference plasmid of genetically modified rapeseed Ms8

YU Xiaobo¹, SHA Yuebing¹, ZHANG Jiangdong¹, ZHAO Lei¹, CHEN Yi¹, SUI Zhiwei²

1.Zhejiang Province Institute of Metrology, Hangzhou 310018, China;
2. National Institute of Metrology, Beijing 100013, China

Received:2016-12-29 Online:2017-05-20 Published:2017-06-06

摘要/Abstract

摘要： 随着转基因产品的大规模商业化和我国大规模进口油菜籽,有效管控存在的风险显得非常必要和有意义,试验研究了质粒标准分子pMs8的研制过程,包括质粒分子的构建、纯度分析、特异性检测、均匀性检验、稳定性考察、定值。结果表明,该质粒分子标准物质的内源基因和外源基因的均匀性良好,没有扩增出所选的其他特异性基因,短期稳定性在14 d(处理温度大于25 ℃)发生显著变化,长期稳定性大于6个月,两种方法对外源基因拷贝数的定值结果相近,因此,pMs8是有准确量值的质粒标准分子。可替代性研究结果表明,其可替代基因组作为转基因油菜Ms8定量的阳性标准。

关键词: 转基因油菜, 数字PCR, 质粒标准分子, Ms8

Abstract: With the commercialization of genetically modified organism (GMO) and large scale of rapeseed imported to China, it was essential to manage and control existing risks. In this paper, genetically modified rapeseed Ms8 was prepared including analysis on preparation, purity, homogeneity, stability and certified value. The results showed that the homogeneity of endogenous gene and exogenous gene of reference molecule was well. No other selective specific gene had been amplified by PCR. Short term stability of reference molecule was obviously changed which were stored at room temperature for 14 days, and long term stability of reference molecule was more than 6 months. The certified value of exogenous gene of reference molecule determined by microspectrophotometric method and digital PCR method was similar. Thus, pMs8 can be used as reference material, and the reference molecule is suitable for the rapid identification of modified rapeseed Ms8 in unknown samples.

Key words: transgenic rapeseed, digital PCR, reference plasmid, Ms8

中图分类号:

S634.3

余笑波, 沙跃兵, 张江东, 赵雷, 陈怡, 隋志伟. 转基因油菜Ms8质粒分子的制备[J]. 浙江农业学报, 2017, 29(5): 694-700.

YU Xiaobo, SHA Yuebing, ZHANG Jiangdong, ZHAO Lei, CHEN Yi, SUI Zhiwei. Development of a reference plasmid of genetically modified rapeseed Ms8[J]. , 2017, 29(5): 694-700.

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转基因油菜Ms8质粒分子的制备

Development of a reference plasmid of genetically modified rapeseed Ms8

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