Isolation and PCR detection of foreign DNA sequences in bee honey raised on genetically modified Bt (Cry1Ac) cotton
Isolation and PCR detection of foreign DNA sequences in bee honey raised on genetically modified Bt (Cry1Ac) cotton
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DOI：
10.1205/fbp06056
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发表时间：
2007-06-01
影响因子：
4.6
通讯作者：
Jia, S.
中科院分区：
文献类型：
--
作者：
Cheng, H.;Jin, W.;Jia, S.
从1997年开始，转基因抗虫棉在中国开始大规模种植。2004年棉花种植面积达370万公顷，占棉花种植总面积的66%。由于棉花是主要的蜜源植物之一，在国际贸易中是否含有外源DNA序列引起了国际贸易的关注。因此，建立与Bt棉相关的蜂蜜检测方法已成为当务之急。我们建立了一种检测蜂蜜中外源DNA序列的DNA提取方法和一种聚合酶链式反应方法。描述了从蜂蜜中提取DNA的详细步骤。从蜂蜜中提纯DNA的一个关键步骤是用缓冲能力较强的磷酸盐缓冲盐水(PBS)分离出高含量的糖。利用聚合酶链式反应技术，可以特异扩增125-550bp的外源DNA序列和棉花内源DNA序列。
Genetically modified Bt cotton has been planted in large scale in China since 1997. Its acreage reached 3.7 million ha, accounting for 66% of the total cotton cultivation area in 2004. Since cotton is one of the major honey-source plants, it raises a concern in the international trade whether there are any alien DNA sequences contained in honey. Therefore, development of detection methods for bee honey associated with Bt cotton has become urgently needed. We have developed a DNA extraction procedure and a PCR protocol for the detection of foreign DNA sequences in bee honey. A detailed step-by-step procedure for the extraction of DNA from honey was described. A key step for purifying DNA from bee honey is to separate out high amount of sugars by using phosphate buffered saline (PBS) with stronger buffering capacity. With PCR protocol, alien and cotton endogenous DNA sequences with a length of 125-550 bp can be specifically amplified.