Intergeneric conjugation in holomycin-producing marine Streptomyces sp strain M095 | |
Hou, Yan-Hua1,2,3,4; Li, Fu-Chao1; Wang, Shu-Jun4; Qin, Song1; Wang, Quan-Fu3,4 | |
刊名 | MICROBIOLOGICAL RESEARCH
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2008 | |
卷号 | 163期号:1页码:96-104 |
关键词 | Genetic Transformation Holomycin Intergeneric Conjugation Marine Streptomyces Protoplast |
ISSN号 | 0944-5013 |
DOI | 10.1016/j.micres.2006.07.003 |
文献子类 | Article |
英文摘要 | Marine Streptomyces are potential candidates for novel natural products and industrial catalysts. In order to set up biosynthesis approach for a holomycin-producing strain M095 isotated from Jiaozhou Bay, China, a genetic transformation system was established using intergeneric conjugation. The plasmid pIJ8600 consists of an origin of replication for Escherichia coli, a phage integrase directing efficient site-specific integration in bacterial chromosome, thiostrepton-induced promoter and an attP sequence. Using E. coli ET12567 (pUZ8002) carrying pIJ8600 as a conjugal donor, while it was mated with strain M095, pIJ8600 was mobilized to the recipient and the transferred DNA was also integrated into the recipient chromosome. The frequency of exconjugants was 1.9 +/- 0.13 x 10(-4) per recipient cell. Analysis of eight exconjugants showed pIJ8600 was stable integrated at a single chromosomal site (attB) of the Streptomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of growth and antimicrobial activity analysis indicated that the integration of pIJ8600 did not seem to affect the biosynthesis of antibiotics or other essential amino acids. To demonstrate the feasibility of above gene transfer system, the allophycocyanin gene (apc) from cyanobacterium Anacystis nidulans UTEX625 was expressed in strain M095, and the results indicated heterologous allophycocyanin could be expressed and folded effectively. (c) 2006 Elsevier GmbH. All rights reserved.; Marine Streptomyces are potential candidates for novel natural products and industrial catalysts. In order to set up biosynthesis approach for a holomycin-producing strain M095 isotated from Jiaozhou Bay, China, a genetic transformation system was established using intergeneric conjugation. The plasmid pIJ8600 consists of an origin of replication for Escherichia coli, a phage integrase directing efficient site-specific integration in bacterial chromosome, thiostrepton-induced promoter and an attP sequence. Using E. coli ET12567 (pUZ8002) carrying pIJ8600 as a conjugal donor, while it was mated with strain M095, pIJ8600 was mobilized to the recipient and the transferred DNA was also integrated into the recipient chromosome. The frequency of exconjugants was 1.9 +/- 0.13 x 10(-4) per recipient cell. Analysis of eight exconjugants showed pIJ8600 was stable integrated at a single chromosomal site (attB) of the Streptomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of growth and antimicrobial activity analysis indicated that the integration of pIJ8600 did not seem to affect the biosynthesis of antibiotics or other essential amino acids. To demonstrate the feasibility of above gene transfer system, the allophycocyanin gene (apc) from cyanobacterium Anacystis nidulans UTEX625 was expressed in strain M095, and the results indicated heterologous allophycocyanin could be expressed and folded effectively. (c) 2006 Elsevier GmbH. All rights reserved. |
学科主题 | Microbiology |
URL标识 | 查看原文 |
语种 | 英语 |
WOS记录号 | WOS:000252803200011 |
公开日期 | 2010-12-24 |
内容类型 | 期刊论文 |
源URL | [http://ir.qdio.ac.cn/handle/337002/5567] ![]() |
专题 | 海洋研究所_实验海洋生物学重点实验室 |
作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China 3.Harbin Inst Technol, Sch Ocean, Weihai 264209, Peoples R China 4.Huaihai Inst Technol, Jiangsu Key Lab Marine Biotechnol, Jiangsu, Peoples R China |
推荐引用方式 GB/T 7714 | Hou, Yan-Hua,Li, Fu-Chao,Wang, Shu-Jun,et al. Intergeneric conjugation in holomycin-producing marine Streptomyces sp strain M095[J]. MICROBIOLOGICAL RESEARCH,2008,163(1):96-104. |
APA | Hou, Yan-Hua,Li, Fu-Chao,Wang, Shu-Jun,Qin, Song,&Wang, Quan-Fu.(2008).Intergeneric conjugation in holomycin-producing marine Streptomyces sp strain M095.MICROBIOLOGICAL RESEARCH,163(1),96-104. |
MLA | Hou, Yan-Hua,et al."Intergeneric conjugation in holomycin-producing marine Streptomyces sp strain M095".MICROBIOLOGICAL RESEARCH 163.1(2008):96-104. |
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