天花粉蛋白基因导入大豆的研究

A Study on Transformation of TCS Gene to Soybean

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。

In order to obtain transgenic soybean plants,which are resistant to diseases and insects,the ti-chosanthin（ TCS） gene was transformed into soybean plants.First the plant expression vector pC-tPro-TCS-GUS was constructed,then transformed into E.coli DH5α.SDS-PAGE results showed that TCS gene could be ex-pressed in E.coli.TCS gene was transformed into the cotyledonary node of soybean Hefeng35 througha grobac-terium-mediated method.The conversion rate was 1.033%with 19 transgenic plants obtained in the T0 genera-tion.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 andT1 generations.