利用农杆菌侵染新技术在豌豆中瞬时表达haFGF.pdfVIP

生物技术通报 ·研究报告· BIOTECHNOLOGY BULLETIN 2013年第10期 利用农杆菌侵染新技术在豌豆中瞬时表达 ha FGF 杨丽萍  金太成  周晓馥 （吉林师范大学生命科学学院，四平 136000） 摘 要 ： 采用一种操作简单的侵染方法，通过豌豆种子吸收携带病毒载体PEBV 的农杆菌菌液进行ha FGF 蛋白的表达，并 采用半定量RT-PCR、Western blotting 和考马斯亮蓝法对蛋白表达的结果进行定量分析。结果表明，利用来源于PEBV 的病毒载体 在豌豆 （ L. ）中成功表达了GFP 和ha FGF 蛋白，对菌液浓度等条件的优化显著提高了GFP 的表达效率，植物中药 Pisum sativum 用蛋白ha FGF 的表达水平占总可溶性蛋白的2.6%。该瞬时表达体系具有独特优势，如简便、高效、表达稳定，在生产药用蛋白 方面具有广阔的应用前景。 关键词 ： 人酸性成纤维细胞生长因子 种子吸收法 豌豆早褐病毒 绿色荧光蛋白 Transient Expression of ha FGF in Pea Plants by New Agroinoculation Biotechnology Yang Liping Jin Taicheng Zhou Xiaofu （ ， ， 136000 ） The School of Life Science Jilin Normal University Siping Abstract:  The paper described a simple system to product ha FGF protein in pea plants by seed absorbing the Agrobacterium suspension containing PEBV-based vectors, and the expression results was detected by semi-quantitative RT-PCR, Western blotting, and Coomassie brilliant blue method. It has been shown that GFP and ha FGF protein were successfully expressed in pea （ L. ）plants. The optimization Pisum sativum of Agrobacterium cell density dramatically improved expression efficiency of GFP protein. The amount of recombinant pharmaceutical ha FGF in plants reached up to 2.6% of the total soluble proteins. The new system has the special advantages as easiest way, high efficiency, stable expression, and has the wide application in production of pharmaceutical proteins in the future. Key words:  Human acidic fi