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天花粉蛋白基因导入大豆的研究
A Study on Transformation of TCS Gene to Soybean
【摘要】 为获得含有天花粉蛋白(TCS)的转基因大豆植株,提高大豆的抗病虫能力,进行了植物表达载体的构建及转入大豆的研究。首先构建植物表达载体p C-t Pro-TCS-GUS,将其转化E.coli DH5α,经SDS-PAGE结果表明:克隆得到的TCS基因可以在大肠杆菌中表达。通过根癌农杆菌介导法,将TCS基因导入大豆合丰35中,获得了19株T0代转基因大豆,转化率为1.033%。选取T0代种子种植在大田中共获得18株T1代转基因大豆,对获得的T0代和T1代转基因植株进行PCR和PCR-Southern检测,证实外源天花粉蛋白基因己经整合到大豆基因组中。
【Abstract】 In order to obtain transgenic soybean plants,which are resistant to diseases and insects,the tichosanthin( TCS) gene was transformed into soybean plants.First the plant expression vector p C-t Pro-TCS-GUS was constructed,then transformed into E. coli DH5α. SDS-PAGE results showed that TCS gene could be expressed in E.coli.TCS gene was transformed into the cotyledonary node of soybean Hefeng 35 through agrobacterium-mediated method.The conversion rate was 1.033% with 19 transgenic plants obtained in the T0 generation.Then 18 strains in the T1 generation were harvested after selecting T0 generation seeds which were planted in the field.PCR and PCR-Southern analysis proved that the TCS gene was integrated into the soybean genome both in the T0 and T1generations.
【Key words】 soybean; TCS gene; vector construction; transgenic plants;
- 【文献出处】 江西农业大学学报 ,Acta Agriculturae Universitatis Jiangxiensis , 编辑部邮箱 ,2015年04期
- 【分类号】S565.1
- 【下载频次】107