Expression and self-assembly of Heterocapsa circularisquama RNA virus-like particles synthesized in Pichia pastoris
Expression and self-assembly of Heterocapsa circularisquama RNA virus-like particles synthesized in Pichia pastoris作者机构：Department of Chemical and Biochemical Engineering Chosun University Gwangju 501-759 Republic of Korea Department of Environmental Engineering Chosun University Gwangju 501-759 Republic of Korea Seoul Center Korea Basic Science Institute Seoul 136-713 Republic of Korea Biotechnology Center of Shandong Academy of Sciences Jinan 250014 China
出 版 物：《Chinese Science Bulletin》
年 卷 期：2012年第57卷第25期
页 面：3288-3293页
核心收录：
学科分类：0710[理学-生物学] 1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 07[理学] 071005[理学-微生物学] 10[医学]
基 金：supported by the Pioneer Research Center Program through the National Research Program of Korea funded by the Ministry of Education Science and Technology Korea (Grant M1071118001-08M1118-00110)(2010)
主 题：巴斯德毕赤酵母 病毒样颗粒 RNA病毒 pastoris 基因合成 自组装 透射电子显微镜分析 紫外分光光度法
摘 要：Heterocapsa circularisquama RNA virus(HcRNAV) is the first single-stranded RNA virus to be characterized that infects *** ability of HcRNAV coat protein(HcRNAV CP) to self-assemble into virus-like particles(VLPs) in vitro suggested that heterologous expression was possible,and that the VLPs might be ideal nanocontainers for the targeted delivery of genes and *** this paper,we report the expression of a codon-optimized HcRNAV 109 CP gene in Pichia pastoris and the production of self-assembled HcRNAV VLPs using large-scale *** HcRNAV 109 CP gene was synthesized according to the codon preference of *** and cloned into a pPICZA *** recombinant plasmid pPICZA-CPsyns was transformed into *** by *** resulting yeast colonies were screened by PCR and analyzed for protein expression by SDS polyacrylamide gel *** large-scale fermentation,the yield of HcRNAV CPsyns reached approximately 2.5 g L 1 within 4 *** HcRNAV VLPs were purified using PEG precipitation followed by cesium chloride density gradient ultracentrifugation,and were subsequently analyzed using UV spectrophotometry and transmission electron *** dye-labeled myoglobin was loaded into the cages of the HcRNAV VLPs and the encapsulation was confirmed by fluorescence *** results point to the possible utilization in pharmacology or nanotechnology of HcRNAV VLPs produced by *** fermentation.
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