萝卜抗真菌蛋白基因Rs-AFPs在大肠杆菌中的表达及其转化番茄的研究

 通过基因工程手段将萝卜抗真菌蛋白基因Rs-AFP_{1( 2)} 插入大肠杆菌表达载体pTrxFus中并诱导表达, 其融合表达产物约22 kD, 约占总可溶蛋白的0. 45% ( 0. 5%) 。抑菌活性试验表明: 重组Rs-AFP_{1( 2)}有抑菌活性, 其中Rs􀀁AFP2 较Rs􀀁AFP1 抑菌活性强。构建了Rs-AFP₂ 基因的植物表达载体pBIAFP₂。利用农杆菌介导法将pBIAFP₂ 导入番茄中, 并诱导出完整的植株32 株。对其中28 株进行PCR 和PCR-Southern Blot 检测, 其中17 株呈阳性。对经PCR Southern Blot 检测呈阳性的植株进行Southern Blot 检测, 6 株呈阳性。

The Expression of Raphnus sativus-Antifungal Protein Gene and Its Transformation into Tomato

c cysteine rich,antifungal proteins.The Rs AFP 1(2) genes were inserted into the vector pTrxFus and transferred into E.coli GI724.The fusion protein from recombinant pTrxAFP 1(2) was about 22kD and taking up 0.4 and 0.5 percent of the total proteins respectively.Inhibition experiment showed that the expressed products had anti fungal activity to Colletotrichum musae.Expressed products from pTrxAFP 2 showed much stronger inhibitory activity than that from pTrxAFP 1.A plant expression vector pBIAFP 2 was constructed, which had been transferred into tomato by Agrobactrium mediated transformation.32 plantlets had been obtained and 28 of them detected by PCR and PCR Southern Blot.The results showed that 17 of them were positive,and the results of Southern Blot showed 6 of them were positive.