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植物学报  2012 

大麦黄矮病毒运动蛋白是RNA沉默抑制因子

DOI: 10.3724/SP.J.1259.2012.00395, PP. 395-404

Keywords: 大麦黄矮病毒,运动蛋白,RNA沉默,抑制因子

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Abstract:

?大麦黄矮病毒(barleyyellowdwarfvirus,BYDV)属黄症病毒科家族,其基因组包含6个开放阅读框(openreadingframes,ORFs)。将BYDV的6个基因分别克隆到pWEIMING101载体上,得到重组基因。电击转化农杆菌后,利用农杆菌瞬时表达方法渗透注射转GFP基因的本氏烟草16c植株的叶片,在长波长紫外灯下观察GFP的表达,并通过Northernblot证明所得现象。研究结果表明,BYDV的PAV株系ORF4编码的运动蛋白(movementprotein,MP)是RNA沉默抑制因子,其表达可以抑制局部和系统RNA沉默。BYDV-MP与GFP的双链RNA(dsGFP)共表达后仍能抑制RNA沉默,荧光强度与叶片中GFP的mRNA和其沉默降解形成的siRNA的量有对应关系,其N端核定位序列对抑制局部基因沉默起主要作用,第5、6位氨基酸是抑制基因沉默的关键氨基酸。BYDV-MP单独渗透注射的部位均产生细胞死亡。

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