{"title":"基因产物(干扰素)在酵母中表达的异质性分析。","authors":"H Wang, B Gao, P Chen, L Dong, Y Li","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>FPLC, SDS-PAGE and Western blot techniques are used to analyse the heterogeneity of interferon alpha A (IFN-alpha A) expressed in yeast. The heterogeneity consists of (i) the presence of IFN polymer, (ii) partial processing of signal leader peptide and (iii) internal degradation. The reasons for heterogeneity of gene products in expression system of yeast are analysed. The methods of avoiding heterogeneity, such as depolymerization, adding inhibitors of protease to the culture supernatant, the oligonucleotide-directed deletion mutagenesis and improvements of fermentation, are discussed.</p>","PeriodicalId":21648,"journal":{"name":"Science in China. Series B, Chemistry, life sciences & earth sciences","volume":"38 8","pages":"954-62"},"PeriodicalIF":0.0000,"publicationDate":"1995-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Analysis of heterogeneity of gene products (interferon) expressed in yeast.\",\"authors\":\"H Wang, B Gao, P Chen, L Dong, Y Li\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>FPLC, SDS-PAGE and Western blot techniques are used to analyse the heterogeneity of interferon alpha A (IFN-alpha A) expressed in yeast. The heterogeneity consists of (i) the presence of IFN polymer, (ii) partial processing of signal leader peptide and (iii) internal degradation. The reasons for heterogeneity of gene products in expression system of yeast are analysed. The methods of avoiding heterogeneity, such as depolymerization, adding inhibitors of protease to the culture supernatant, the oligonucleotide-directed deletion mutagenesis and improvements of fermentation, are discussed.</p>\",\"PeriodicalId\":21648,\"journal\":{\"name\":\"Science in China. Series B, Chemistry, life sciences & earth sciences\",\"volume\":\"38 8\",\"pages\":\"954-62\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1995-08-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Science in China. Series B, Chemistry, life sciences & earth sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Science in China. Series B, Chemistry, life sciences & earth sciences","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
采用FPLC、SDS-PAGE和Western blot技术分析干扰素α A (ifn - α A)在酵母中表达的异质性。非均质性包括(i) IFN聚合物的存在,(ii)信号先导肽的部分加工和(iii)内部降解。分析了酵母表达系统中基因产物异质性的原因。讨论了解聚、在培养上清液中加入蛋白酶抑制剂、寡核苷酸定向缺失诱变和改进发酵等避免非均质性的方法。
Analysis of heterogeneity of gene products (interferon) expressed in yeast.
FPLC, SDS-PAGE and Western blot techniques are used to analyse the heterogeneity of interferon alpha A (IFN-alpha A) expressed in yeast. The heterogeneity consists of (i) the presence of IFN polymer, (ii) partial processing of signal leader peptide and (iii) internal degradation. The reasons for heterogeneity of gene products in expression system of yeast are analysed. The methods of avoiding heterogeneity, such as depolymerization, adding inhibitors of protease to the culture supernatant, the oligonucleotide-directed deletion mutagenesis and improvements of fermentation, are discussed.