利用浸苗法将野生天麻总DNA导入马铃薯的研究

目的研究野生天麻总DNA对马铃薯的转化,分析马铃薯转化植株中野生天麻的药用成分天麻素。方法采用浸苗法将野生天麻总DNA导入马铃薯试管苗,通过紫外扫描法、PCR扩增筛选转化植株,对转化植株进行SDS-聚丙烯酰胺凝胶电泳(SDS—PAGE)蛋白分析,通过TLC法检测天麻素。结果(1)在200株转化的马铃薯中有21株的紫外扫描图谱与正常对照组有显著差异,且在220 nm有明显吸收峰。(2)5株经PCR扩增出野生天麻抗真菌蛋白(GAFP)基因。(3)转基因马铃薯与正常马铃薯的蛋白表达有明显差异,并且在转基因马铃薯中有一条与GAFP相同的条带。而正常马铃薯中无此条带。(4)通过薄层色谱法检测出3株转基因马铃薯表达野生天麻的有效药用成分天麻素。结论采用浸苗法进行外源总DNA导入是可行的。

A genetic transformation study of introducing wild Gastrodia elata DNA into Solanum tuberosum by soaking seedling method

Objective Using the soaking seedling method to introduce wild Gastrodia elata DNA into potato plantlets and analyze gastrodin of transformed Solanum tuberosum.Methods After the tuber grown up,the solution of gastrodin was extracted from the potatoes which were transformed by wild G.(elata) DNA.The transformed plants were scaned by ultraviolet.PCR was used to analyze GAFP gene by SDS-PAGE.TLC was used to analyze the gastrodin of transformed S.tuberosum.Results(1)The 21 transformed plants had obvious absorption peak at 220 nm in 200 transformed S.tuberosunr.(2)The fragment of wild G.elata GAFP gene was found by PCR from 5 transformed plants.(3)The protein expressions in transgenic S.tuberosum were obviously different from the normal S.tuberosum;a band was detected in transgenic S.tuberosum and wild G.elata,but wasn't detected in normal S.tuberosum.(4)The expression of the gastrodin was detected by TLC from three transformed plants. Conclusion It is feasible to use the soaking seedling method to introduce exogenous DNA into plants.