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抑制南方水稻黑条矮缩病毒非结构蛋白P5-1的表达阻碍病毒在昆虫体内的增殖

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Keywords: 南方水稻黑条矮缩病毒,P5-1蛋白,抗体制备,RNA干扰,增殖

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Abstract:

为明确南方水稻黑条矮缩病毒(Southernriceblack-streakeddwarfvirus,SRBSDV)编码的非结构蛋白P5-1参于SRBSDV在介体白背飞虱体内侵染过程中的作用机制,通过原核表达蛋白制备SRBSDV编码的非结构蛋白P5-1的多克隆抗体,并应用Westernblot和免疫荧光标记法检测抗体的特异性,以注射法将来源于P5-1基因的dsRNA(dsP5-1)注入获毒1d的白背飞虱体内,5d后通过免疫荧光标记法检测dsP5-1对SRBSDV在白背飞虱体内增殖的影响,同时以注射来源于GFP基因的dsRNA(dsGFP)为对照.结果显示,Westernblot和免疫荧光标记分别检测到SRBSDV侵染水稻和白背飞虱表达的P5-1蛋白,表明所制备的P5-1抗体具有特异性.dsGFP处理的对照组白背飞虱带毒率高达81%,而dsP5-1处理的白背飞虱带毒率仅为21%,且P5-1蛋白的表达和SRBSDV在昆虫体内的增殖均受到抑制.表明P5-1蛋白是病毒在昆虫体内增殖的关键因子,可作为阻断病毒在昆虫体内增殖的理想靶标.

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