Sudarshana Mysore R, Plesha Michael A, Uratsu Sandra L, Falk Bryce W, Dandekar Abhaya M, Huang Ting-Kuo, McDonald Karen A
Department of Plant Pathology, University of California at Davis, One Shields Ave., Davis, CA 95616, USA.
Plant Biotechnol J. 2006 Sep;4(5):551-9. doi: 10.1111/j.1467-7652.2006.00202.x.
A novel cucumber mosaic virus inducible viral amplicon (CMViva) expression system has been developed that allows for tightly regulated chemically inducible expression of heterologous genes in plant hosts. Transient production of recombinant alpha(1)-antitrypsin (rAAT), a human blood protein, was demonstrated in Nicotiana benthamiana leaves. The highest production levels were obtained by co-infiltrating leaves with Agrobacterium tumefaciens cells containing CMViva carrying the AAT gene and A. tumefaciens cells carrying a binary vector constitutively expressing the gene silencing suppressor p19. Accumulation of up to thirty-fold more rAAT was observed in leaves (24 mg per 100 g leaf tissue) when compared with the expression levels observed using the cauliflower mosaic virus (CaMV) 35S promoter. Significantly, 70% of the rAAT produced using the CMViva expression system was found to be biologically active, a 170-fold increase in functional protein compared with the CaMV 35S expression system.
已开发出一种新型黄瓜花叶病毒诱导型病毒扩增子（CMViva）表达系统，该系统可在植物宿主中对异源基因进行严格调控的化学诱导表达。在本氏烟草叶片中证实了人血蛋白重组α1-抗胰蛋白酶（rAAT）的瞬时产生。通过将携带AAT基因的CMViva的根癌农杆菌细胞与携带组成型表达基因沉默抑制子p19的二元载体的根癌农杆菌细胞共同浸润叶片，获得了最高产量水平。与使用花椰菜花叶病毒（CaMV）35S启动子观察到的表达水平相比，在叶片中观察到rAAT积累量增加了多达30倍（每100克叶片组织24毫克）。值得注意的是，发现使用CMViva表达系统产生的rAAT中有70%具有生物活性，与CaMV 35S表达系统相比，功能蛋白增加了170倍。
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