A. Rostami, N. S. Bashir, Parviz Pirniakan, N. Masoudi
{"title":"黄瓜花叶病毒外壳蛋白的表达及其组装成病毒样颗粒","authors":"A. Rostami, N. S. Bashir, Parviz Pirniakan, N. Masoudi","doi":"10.17795/BHS-24729","DOIUrl":null,"url":null,"abstract":"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.","PeriodicalId":8849,"journal":{"name":"Biotechnology and Health Sciences","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2014-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"7","resultStr":"{\"title\":\"Expression of Cucumber Mosaic Virus Coat Protein and Its Assembly into Virus Like Particles\",\"authors\":\"A. Rostami, N. S. Bashir, Parviz Pirniakan, N. Masoudi\",\"doi\":\"10.17795/BHS-24729\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"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.\",\"PeriodicalId\":8849,\"journal\":{\"name\":\"Biotechnology and Health Sciences\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2014-11-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"7\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biotechnology and Health Sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.17795/BHS-24729\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology and Health Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.17795/BHS-24729","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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.