Hao Li, Ling-Yan Wang, Gui-Yun Xu, Yang Chen, Rong Jiang, Yuan Li
{"title":"链霉菌udp -葡萄糖脱氢酶编码基因Ste6的克隆、表达和特性研究","authors":"Hao Li, Ling-Yan Wang, Gui-Yun Xu, Yang Chen, Rong Jiang, Yuan Li","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>Streptomyces sp. 139 was identified to produce a new exopolysaccharide Ebosin (139A) with antirheumatic arthritis activity in vivo. The Ebosin biosynthesis gene cluster(31.3 kb; GenBank Accession Number: AY131229) containing 22 ORFs (ste1-ste22) of Streptomyces sp. 139 had been reported previously. In this paper,we present experimental evidence for the identity of the ste6 gene product as a UDP-glucose dehydrogenase (UDPGDH). With pET-30a as vector,the gene was cloned and expressed in Escherichia coli BL21 (DE3). The expressed protein was purified to homogeneity by His-Bind resin affinity chromatograpy and it was able to catalyze UDP-glucose to UDP-glucuronic acid. To evaluate the function of ste6, the gene was disrupted by a single-crossover homologous recombination event and the result showed that ste6 is required in Ebosin biosynthesis.</p>","PeriodicalId":23770,"journal":{"name":"Yi chuan xue bao = Acta genetica Sinica","volume":"32 11","pages":"1213-20"},"PeriodicalIF":0.0000,"publicationDate":"2005-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Cloning, expression and characterization of the Ste6 gene encoding a UDP-glucose dehydrogenase in Streptomyces.\",\"authors\":\"Hao Li, Ling-Yan Wang, Gui-Yun Xu, Yang Chen, Rong Jiang, Yuan Li\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Streptomyces sp. 139 was identified to produce a new exopolysaccharide Ebosin (139A) with antirheumatic arthritis activity in vivo. The Ebosin biosynthesis gene cluster(31.3 kb; GenBank Accession Number: AY131229) containing 22 ORFs (ste1-ste22) of Streptomyces sp. 139 had been reported previously. In this paper,we present experimental evidence for the identity of the ste6 gene product as a UDP-glucose dehydrogenase (UDPGDH). With pET-30a as vector,the gene was cloned and expressed in Escherichia coli BL21 (DE3). The expressed protein was purified to homogeneity by His-Bind resin affinity chromatograpy and it was able to catalyze UDP-glucose to UDP-glucuronic acid. To evaluate the function of ste6, the gene was disrupted by a single-crossover homologous recombination event and the result showed that ste6 is required in Ebosin biosynthesis.</p>\",\"PeriodicalId\":23770,\"journal\":{\"name\":\"Yi chuan xue bao = Acta genetica Sinica\",\"volume\":\"32 11\",\"pages\":\"1213-20\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2005-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Yi chuan xue bao = Acta genetica Sinica\",\"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":"Yi chuan xue bao = Acta genetica Sinica","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Cloning, expression and characterization of the Ste6 gene encoding a UDP-glucose dehydrogenase in Streptomyces.
Streptomyces sp. 139 was identified to produce a new exopolysaccharide Ebosin (139A) with antirheumatic arthritis activity in vivo. The Ebosin biosynthesis gene cluster(31.3 kb; GenBank Accession Number: AY131229) containing 22 ORFs (ste1-ste22) of Streptomyces sp. 139 had been reported previously. In this paper,we present experimental evidence for the identity of the ste6 gene product as a UDP-glucose dehydrogenase (UDPGDH). With pET-30a as vector,the gene was cloned and expressed in Escherichia coli BL21 (DE3). The expressed protein was purified to homogeneity by His-Bind resin affinity chromatograpy and it was able to catalyze UDP-glucose to UDP-glucuronic acid. To evaluate the function of ste6, the gene was disrupted by a single-crossover homologous recombination event and the result showed that ste6 is required in Ebosin biosynthesis.