摘要
从黄瓜花叶病毒(CMV)中提取总RNA,通过反转-录PCR(RT-PCR)扩增得到其运动蛋白2 b基因,将扩增产物克隆到pMD-18T载体上。DNA序列分析表明,所得到2 b基因全长为303 bp与已发表CMV序列相同。将目的片段亚克隆到表达载体pET-30 a上,并在大肠杆菌JM109诱导表达,诱导9 h后,融合蛋白表达量最大。经SDS-PAGE检测,融合蛋白以可溶形式存在。制备的抗血清,效价为1:4 000。
Cloning 2 b protein gene of cucumber mosiac virus, the movement protein gene obtained by RT-PCR from the total RNA of cucumber mosaic virus was cloned to pMD-18T vector. The target fragment was subcloned to pET-30 a, an expression vector. The fusion protein was expressed in the stain JM109 of Eseherichia coll. Its length of 2 b gene was 303 bp. The highest expression amount of the fusion protein, which was soluble after sonication, was 9 hr after inducing. The titer of target fusion protein antiserum was 1：4000.
出处
《药物生物技术》
CAS
CSCD
2006年第1期6-8,共3页
Pharmaceutical Biotechnology
关键词
黄瓜花叶病毒
运动蛋白
抗血清
融合蛋白表达
Cucumber mosaic virus, Movement protein, cDNA, Fusion protein expression
