[Detection of recombinant DNA from genetically modified papaya].
[Detection of recombinant DNA from genetically modified papaya].
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转基因木瓜重组DNA的检测[J].
DOI：
10.3358/shokueishi.42.231
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
M. Toyoda
中科院分区：
文献类型：
--
作者：
Y. Goda;T. Asano;M. Shibuya;A. Hino;M. Toyoda
建立了一种检测转基因番木瓜（55-1系）的聚合酶链反应（PCR）方法。木瓜内源木瓜蛋白酶基因被用作内部对照。木瓜蛋白酶基因片段的PCR扩增结果表明，商业硅胶膜型试剂盒（QIAGEN DNeasy plant mini）可用于从木瓜果实中提取DNA，但不适用于从木瓜罐头中提取DNA。另一方面，商业离子交换型试剂盒（QIAGEN Genomic-tip）提供了足够的纯化DNA用于罐装木瓜果实的PCR。结果表明，与亲本及其它非转基因番木瓜品种相比，转基因番木瓜果实DNA在5对引物（Nos. 2-6)包括β-葡糖醛酸糖苷酶和新霉素磷酸转移酶II基因特异性的那些。另一方面，当我们使用来自罐装木瓜的DNA时，识别这些基因的引物对显示假阳性结果。因此，我们建议引物对（编号：5和6）识别来自两种不同生物的序列，以特异性地检测罐头水果中的转基因木瓜。
A method using polymerase chain reaction (PCR) was developed to detect the genetically modified (GM) papaya (55-1 line), of which the mandatory safety assessment has not been finished in Japan because of insufficient data. The papaya intrinsic papain gene was used as an internal control. The results of PCR amplification of the papain gene segment indicated that a commercial silica membrane type kit (QIAGEN DNeasy plant mini) was useful for extraction of DNA from papaya fruit, but not for extraction from canned papaya fruit. On the other hand, a commercial ion-exchange type kit (QIAGEN Genomic-tip) provided enough purified DNA for PCR from canned papaya fruit. Compared with the parental line and other commercial non-GM papayas, the DNA from GM papaya fruit provided specific amplification bands in PCR with five primer pairs (Nos. 2-6) including beta-glucuronidase and neomycin phosphotransferase II gene-specific ones. On the other hand, the primer pairs recognizing these genes showed false-positive results when we used DNAs from canned papaya. Therefore, we recommend that the primer pairs (Nos. 5 and 6) recognizing the sequences derived from two different species of organism should be used in order to detect specifically the GM papaya in canned fruits.