番茄环纹斑点病毒非结构蛋白NSs的原核表达及多抗血清制备
Southwest China Journal of Agricultural Sciences(2015)
Abstract
用RT-PCR从感染番茄环纹斑点病毒(Tomato zorate spot virus,TZSV)的番茄中扩增出编码病毒非结构蛋白NSs基因,将NSs基因构建到原核表达载体pET-30 a中,获得原核表达载体pET-30a-NSs,转化大肠杆菌BL21 (DE3),用IPTG诱导表达.SDS-PAGE电泳分析显示,高效诱导表达了57kDa融合蛋白.用回收纯化的融合蛋白免疫家兔,获得多抗血清.间接ELISA测定多抗血清的效价为1/8192.用制备好的抗血清对TZSV感病病样进行Western blotting检测,能检测到特异性条带.结果表明该抗血清可用于TZSV的检测,并为进一步研究NSs功能奠定了基础.
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