多克隆抗体
生物
抗血清
免疫原
病毒学
重组DNA
分子生物学
病毒
污渍
表达式向量
抗体
单克隆抗体
基因
生物化学
免疫学
作者
Jinha Kim,Ha-Na Yoo,Eun Hui Bae,Yong-Tae Jung
出处
期刊:Journal of Microbiology and Biotechnology
[Journal of Microbiology and Biotechnology]
日期:2012-12-28
卷期号:22 (12): 1776-1781
被引量:12
标识
DOI:10.4014/jmb.1208.08042
摘要
Multiple viruses such as Lily symptomless virus (LSV), Lily mottle virus (LMoV), and cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gangwon, Chungnam, and Jeju provinces of Korea in 2008-2011. Coat protein (CP) genes of LSV and LMoV were amplified from collected samples by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into a pET21d(+) expression vector to generate recombinant CPs. The resulting carboxy-terminal His-tagged CPs were expressed in Escherichia coli strain BL21(DE3) by isopropyl-1-thio- beta-D-galactoside induction. The recombinant proteins were purified using Ni-NTA agarose beads, and the purified proteins were used as an immunogen to produce polyclonal antibodies in rabbits. The resulting polyclonal antisera recognized specifically LSV and LMoV from infected plant tissues in Western blotting assays. Indirect enzyme-linked immunosorbent assay and immunocapture RTPCR using these polyclonal antisera were developed for the sensitive, efficient, economic, and rapid detection of Lily viruses. These results suggest that large-scale bulb tests and economic detection of Lily viruses in epidemiological studies can be performed routinely using these polyclonal antisera.
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