{"title":"研究核糖体RNA折叠辅助蛋白的蛋白质表达管道","authors":"Philipp Vierig, R. Börner","doi":"10.52825/ocp.v2i.162","DOIUrl":null,"url":null,"abstract":"The main focus of this work is to establish a protein synthesis pathway for T7 RNA polymerase (T7-RNAP) and Pumilio and fem3 mRNA binding factors 6 (Puf6) using the minimum necessary methods.The proteins can be cost-effectively expressed pure and highly active in prokaryotes and then effectively purified with a hexahistidine tag via nickel-NTA affinity chromatography. The proteins find application in the synthesis and folding of, for example, ribosomal RNA (rRNA). This paper describes the optimisation of T7 and Puf6 synthesis to ensure the highest possible yield and purity, both indispensable factors for research and biotechnological applications. research and biotechnological application of the proteins.","PeriodicalId":89291,"journal":{"name":"The open conference proceedings journal","volume":"2 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2022-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Protein Expression Pipeline for Research on Folding Helper Proteins for Ribosomal RNA\",\"authors\":\"Philipp Vierig, R. Börner\",\"doi\":\"10.52825/ocp.v2i.162\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The main focus of this work is to establish a protein synthesis pathway for T7 RNA polymerase (T7-RNAP) and Pumilio and fem3 mRNA binding factors 6 (Puf6) using the minimum necessary methods.The proteins can be cost-effectively expressed pure and highly active in prokaryotes and then effectively purified with a hexahistidine tag via nickel-NTA affinity chromatography. The proteins find application in the synthesis and folding of, for example, ribosomal RNA (rRNA). This paper describes the optimisation of T7 and Puf6 synthesis to ensure the highest possible yield and purity, both indispensable factors for research and biotechnological applications. research and biotechnological application of the proteins.\",\"PeriodicalId\":89291,\"journal\":{\"name\":\"The open conference proceedings journal\",\"volume\":\"2 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2022-12-15\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"The open conference proceedings journal\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.52825/ocp.v2i.162\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"The open conference proceedings journal","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.52825/ocp.v2i.162","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Protein Expression Pipeline for Research on Folding Helper Proteins for Ribosomal RNA
The main focus of this work is to establish a protein synthesis pathway for T7 RNA polymerase (T7-RNAP) and Pumilio and fem3 mRNA binding factors 6 (Puf6) using the minimum necessary methods.The proteins can be cost-effectively expressed pure and highly active in prokaryotes and then effectively purified with a hexahistidine tag via nickel-NTA affinity chromatography. The proteins find application in the synthesis and folding of, for example, ribosomal RNA (rRNA). This paper describes the optimisation of T7 and Puf6 synthesis to ensure the highest possible yield and purity, both indispensable factors for research and biotechnological applications. research and biotechnological application of the proteins.