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对在固体培养基上培养3年的黄瓜转gfp基因毛状根进行gfp和rol位点系列基因的PCR扩增、gfp的荧光定量PCR、Western blot杂交以及荧光观察分析。结果显示,由组成型启动子35S驱动的gfp在转录水平上能正常表达,而且能够翻译出编码蛋白;此外,培养3年后的毛状根,能扩增出与毛状根形态构成有关的rol系列基因。本研究结果表明长期培养的毛状根能保持其遗传稳定性,这为利用毛状根长期工厂化生产外源基因表达的蛋白产物和药用植物的活性成分提供了理论依据。
The gfp and rol locus genes were amplified by PCR from cucumber gfp hairy roots cultured on solid medium for 3 years. The gfp fluorescence quantitative PCR, Western blot hybridization and fluorescence observation were performed. The results showed that gfp driven by the constitutive promoter 35S was expressed normally at the transcriptional level and was able to translate the encoded protein. In addition, the hairy roots after 3 years of culture can amplify the morphological features of hairy roots rol series of genes. The results of this study show that long-term cultured hairy roots can maintain their genetic stability, which provides a theoretical basis for the long-term factory production of exogenous gene expression of hairy root products and active ingredients of medicinal plants.