兰州分离物百合无症病毒外壳蛋白的表达、纯化与鉴定。
Expression, purification and characterization of the Lily symptomless virus coat protein from Lanzhou Isolate.
机构信息
Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China.
出版信息
Virol J. 2010 Feb 10;7:34. doi: 10.1186/1743-422X-7-34.
BACKGROUND
Lily symptomless virus (LSV) is widespread in many countries where lily are grown or planted, and causes severe economic losses in terms of quantity and quality of flower and bulb production. To study the structure-function relationship of coat protein (CP) of LSV, to investigate antigenic relationships between coat protein subunits or intact virons, and to prepare specific antibodies against LSV, substantial amounts of CP protein are needed.
RESULTS
Thus, full-length cDNA of LSV coat protein was synthesized and amplified by RT-PCR from RNA isolated from LSV Lanzhou isolate. The extended 33.6 kDa CP was cloned and expressed prokaryoticly and then purified by Ni-ion affinity chromatography. Its identity and antigenicity of recombinant CP were identified on Western-blotting by using the prepared anti-LSV antibodies.
CONCLUSIONS
The results indicate that fusion CP maintains its native antigenicity and specificity, providing a good source of antigen in preparation of LSV related antibodies. Detailed structural analysis of a pure recombinant CP should allow a better understanding of its role in cell attachment and LSV tropism. This investigation to LSV should provide some specific antibodies and aid to development a detection system for LSV diagnostics and epidemiologic surveys.
背景
百合无症状病毒（LSV）广泛存在于许多种植或栽培百合的国家，对花卉和鳞茎的产量和质量造成严重的经济损失。为了研究LSV 外壳蛋白（CP）的结构-功能关系，研究 CP 亚基或完整病毒粒子之间的抗原关系，并制备针对 LSV 的特异性抗体，需要大量的 CP 蛋白。
结果
因此，通过从 LSV 兰州分离株中分离的 RNA，通过 RT-PCR 合成并扩增了 LSV 外壳蛋白的全长 cDNA。将扩展的 33.6 kDa CP 进行克隆和原核表达，然后通过 Ni 离子亲和层析进行纯化。用制备的抗 LSV 抗体通过 Western-blotting 鉴定重组 CP 的身份和抗原性。
结论
结果表明，融合 CP 保持其天然的抗原性和特异性，为制备 LSV 相关抗体提供了良好的抗原来源。对纯重组 CP 的详细结构分析应能更好地理解其在细胞附着和 LSV 嗜性中的作用。对 LSV 的这项研究将提供一些特异性抗体，并有助于开发用于 LSV 诊断和流行病学调查的检测系统。
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