烟曲霉WY-1植酸酶基因的克隆及其在烟草中的表达

Cloning of Aspergillus fumigatus WY-1 Phytase Gene and Expression of Gene in Tobacco

以自行筛选的烟曲霉WY-1为出发菌株,通过PCR方法克隆其植酸酶基因,并构建了含有植物信号肽序列的植酸酶基因植物表达载体,利用农杆菌介导的叶盘法对烟草叶片进行了遗传转化。结果表明,PCR鉴定和Southern杂交分析说明植酸酶基因已整合到烟草的基因组中;RT-PCR检测结果证实了植酸酶基因已得到转录表达;酶活性分析表明功能性的植酸酶已成功表达在烟草中,这是烟曲霉植酸酶基因在烟草中的首次表达。

Taking a phytase-producing strain, Aspergillus fumigatus WY-1 as an original strain, a phytase gene was amplified from genomic DNA of the strain by PCR. A plant expression vector containing the phytase gene with a plant signal peptide was ＂constructed and then introduced into tobaccos by an A. tumefaciens mediated transformation protocol. PCR Screening and Southern blotting analysis demonstrated that the phytase gene had been integrated into the genomic DNA of tobacco, and the determination result of RT-PCR. revealed that the phytase gene had been transcript, and the assay of enzyme activity showed that a functional phytase had been successfully expressed in tobaccos. It is the fn＇st time to express an A.fumigatus phytase gene in tobaccos.