黄瓜花叶病毒外壳蛋白的表达及其组装成病毒样颗粒

A. Rostami, N. S. Bashir, Parviz Pirniakan, N. Masoudi
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引用次数: 7

摘要

目的:报道VLP在表达CMVCP基因的大肠杆菌细胞中的表达和组装。材料与方法:在本研究中,CMV-CP基因从预先制备的克隆载体中释放。然后将CMV-CP连接到表达载体上。测序由韩国Marcrogen公司完成。将重组质粒转移到大肠杆菌Rosetta分离株中。经异丙基硫代半乳糖诱导后,用SDS-PAGE测定表达蛋白的分子量。用NATURE法提取蛋白,观察是否存在cmv样颗粒。结果:用CMV特异性多克隆抗体和偶联物Western blotting检测CMV- cp。采用JEOL 100- cxii型透射电镜,放大100000x倍,加速电压100kv,对产CP克隆提取的蛋白进行研究。结论:CP基因在原核系统中成功表达,并组装成cmv样颗粒。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Expression of Cucumber Mosaic Virus Coat Protein and Its Assembly into Virus Like Particles
Objectives: In this paper, we report the expression and assembly of VLP in cells of E. coli expressing the CMVCP gene. Materials and Methods: In this study, the CMV-CP gene was released from a previously prepared cloning vector. Then, the CMV-CP was ligated into the expression vector. Sequencing was done by Marcrogen, Inc. (South Korea). A recombinant plasmid was transferred to E.coli isolate Rosetta. After inducing by isopropyl thiogalactosides, the molecular weight of the expressed protein was determined by SDS-PAGE. The extraction of proteins was done by NATURE method to see the possible presence of CMV-like particles. Results: CMV-CP was detected by Western blotting by a CMV specific polyclonal antibody and conjugate. The protein extracted from the CP producing clone was studied under a JEOL 100-CXII transmission electron microscope with 100000× magnification at an acceleration voltage of 100 kV. Conclusions: The results showed that the CP gene was expressed in the prokaryotic system successfully and was assembled into the CMVlike particle.
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