外源木聚糖酶基因atx在水稻中的表达

Expression of Exogenous Xylanase Gene (atx) in Transgenic Rice

为通过在转基因植株中表达木聚糖酶来提高木聚糖酶的生产效率,将具有较高热稳定性和催化活性的杂合木聚糖酶基因atx连接到双元表达载体pCAMBIA1301上,成功构建了木聚糖酶植物表达载体atx-Ru3ep-1301。然后以水稻成熟胚的愈伤组织作为转化受体,采用农杆菌介导法将木聚糖酶基因导入水稻(中花11)中。经过潮霉素抗性检测和PCR鉴定,证实目的基因已经整合到转基因水稻基因组中。RT-PCR分析结果显示,外源木聚糖酶基因能够在CaMV 35S启动子的引导下在转基因水稻中正常转录。转基因水稻能够正常生长和繁殖。木聚糖酶活性分析表明,转基因植株最高木聚糖酶活性约为4.37 U/g(鲜叶片)。因此,利用转基因水稻生产木聚糖酶将会是一种经济、有效的方法。

For improving the production efficiency of xylanase by expression of xylanase in transgenic rice plants, a xyla- nase gene （atx） was inserted into the binary expression vector pCAMBIA1301, and the resulting construction with atx was named atx-Ru3ep-1301. The atx-Ru3ep-1301 vector was introduced into rice （Oryza satiwa L. subsp, japonica） variety Zhonghua 11 via Agrobacterium-mediated transformation, by using the rice mature embryo-derived callus as the explants. Both hygromycin resistance detection and PCR amplification confirmed that the target gene had been integrated into the ge- nome of transgenic rice. RTPCR analysis of the total RNA extracted from the fresh leaves of several transgenic lines showed that the introduced xylanase gene could be normally expressed in rice under the control of CaMV 358 promoter. And in the leaves of transgenic rice, xylanase activity was up to 4.37 U/g in fresh leaf. In addition, growth and reproduction of the transgenic rice plants were not affected by the expression of foreign xylanase. Thus, it would be an economic way for xyla- nase production via transgenic rice.