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通过定性PCR技术,对棉籽壳培养基及栽培银耳中的转基因成分进行检测。以棉籽壳培养基及栽培银耳的DNA为模板,设置棉花内源基因Sad1内参对照、转基因棉籽壳阳性对照、非转基因棉籽壳阴性对照和无菌双蒸水空白对照,利用不同的引物分别对CaMV35S启动子、NOS终止子以及Cry1A抗虫基因的特异片段进行PCR扩增及电泳检测。在棉籽壳培养基中可检测出35S、NOS及Cry1A的特异片段,而在棉籽壳栽培的银耳中仅检测出35S和NOS的特异片段,未检出Cry1A的特异片段。研究结果表明,棉籽壳培养基中含有外源转基因成分,并向银耳发生水平转移,而其外源抗虫基因可能未转入银耳中。对棉籽壳栽培银耳中基因水平转移的影响因素和潜在生物安全性进行了讨论。
Through qualitative PCR technique, the transgenic components in cottonseed shell culture medium and cultivated Tremella were detected. The cotton endogenous gene Sad1 reference control, transgenic cottonseed shell positive control, non-transgenic cottonseed shell negative control and sterilized double distilled water blank control were set up using the cottonseed shell culture medium and the cultivated Tremella fuciformis as template. Different primers were used to detect CaMV35S Promoter, NOS terminator and Cry1A insect-resistant gene specific fragments for PCR amplification and electrophoresis. The specific fragments of 35S, NOS and Cry1A were detected in the cottonseed shell culture medium, while only the specific fragments of 35S and NOS were detected in the white fungus cultivated in the cottonseed shell. No specific fragment of Cry1A was detected. The results showed that cottonseed husk culture medium contained foreign genetically modified components and transferred horizontally to white fungus, while the foreign insect-resistant genes may not be transferred to white fungus. The influencing factors and potential biosafety of horizontal gene transfer in cottonseed cultivar Tremella were discussed.