植物中瞬时表达外源基因的新型侵染技术

A new agroinoculation technology for foreign gene expression in plants by means of transient expression

由于瞬时表达系统的局限性使其不利于规模化生产的应用,本文介绍了一种新型的瞬时表达侵染技术—种子吸收法在植物中进行外源蛋白的表达。利用种子自然吸收来源于TMV病毒载体的农杆菌重悬液在番茄中成功的表达了报告基因GFP,并优化了影响基因表达的各种因素,包括菌液重悬液浓度和其他侵染条件。与其它侵染方法相比,该方法具有独特优势,如操作过程简便,有利于外源蛋白的规模化生产,并能够进一步扩大植物生物反应器的宿主范围。我们推测种子吸收法将有利于重组药用蛋白在植物中的工业规模化生产。

Due to the laborious and scale-up limitation we have developed a simple method named ＂seed absorption＂ to express foreign proteins in plants by means of transient expression. It has been shown that the reporter gene GFP was expressed successfully in tomato （Solanum lycopersicum L.） plants by seed absorbing Agrobacterium suspension containing TMV-based p35S-30B-GFP vector. Various factors influencing the gene expression were optimized including Agrobacterium cell density and other inoculation conditions. This method has the special advantages as simple work process, ease to scale-up, and further expanding the host range of plant bioreactor than previous methods. We assume that the seed absorption method will facilitate the industrial scale production of the recombinant pharmaceutical proteins in plants.