attB site disruption in marine Actinomyces sp M048 via DNA transformation of a site-specific integration vector
文献类型：期刊论文
| 作者 | Hou, Yan-Hua1,2; Wang, Quan-Fu2; Ding, Ling1; Li, Fu-Chao1; Qin, Song1 |
| 刊名 | BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
 |
| 出版日期 | 2008-05-01 |
| 卷号 | 50期号:Part 1页码:11-16 |
| 关键词 | Antimicrobial Activity Attb Site Of Actinomyces Strain M048 Bafilomycin Chandrananimycin Dna Transformation Marine Actinomyces |
| ISSN号 | 0885-4513 |
| DOI | 10.1042/BA20070124 |
| 文献子类 | Article |
| 英文摘要 | An efficient conjugation method has been developed for the marine Actinomyces sp. isolate M048 to facilitate the genetic manipulation of the chandrananimycin biosynthesis gene cluster. A phi C31-derived integration vector pIJ8600 containing oriT and attP fragments was introduced into strain M048 by bi-parental conjugation from Escherichia coli ET12567 to strain M048. Transformation efficiency was (6.38 +/- 0.41) x 10(-5) exconjugants per recipient spore. Analysis of eight exconjugants showed that the plasmid pIJ8600 was stably integrated at a single chromosomal site (attB) of the Actinomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of antimicrobial activity analysis indicated that the insertion of plasmid pIJ8600 seemed to affect the biosynthesis of antibiotics that could strongly inhibit the growth of E. coli and Mucor miehei (Tu284). HPLC-MS analysis of the extracts indicated that disruption of the attB site resulted in the complete abolition of chandrananimycin A-C production, proving the identity of the gene cluster. Instead of chandrananimycins, two bafilomycins were produced through disruption of the attB site from the chromosomal DNA of marine Actinomyces sp. M048.; An efficient conjugation method has been developed for the marine Actinomyces sp. isolate M048 to facilitate the genetic manipulation of the chandrananimycin biosynthesis gene cluster. A phi C31-derived integration vector pIJ8600 containing oriT and attP fragments was introduced into strain M048 by bi-parental conjugation from Escherichia coli ET12567 to strain M048. Transformation efficiency was (6.38 +/- 0.41) x 10(-5) exconjugants per recipient spore. Analysis of eight exconjugants showed that the plasmid pIJ8600 was stably integrated at a single chromosomal site (attB) of the Actinomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of antimicrobial activity analysis indicated that the insertion of plasmid pIJ8600 seemed to affect the biosynthesis of antibiotics that could strongly inhibit the growth of E. coli and Mucor miehei (Tu284). HPLC-MS analysis of the extracts indicated that disruption of the attB site resulted in the complete abolition of chandrananimycin A-C production, proving the identity of the gene cluster. Instead of chandrananimycins, two bafilomycins were produced through disruption of the attB site from the chromosomal DNA of marine Actinomyces sp. M048. |
| URL标识 | 查看原文 |
| 语种 | 英语 |
| WOS记录号 | WOS:000255677400002 |
| 公开日期 | 2010-12-30 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/6279]  |
| 专题 | 海洋研究所_实验海洋生物学重点实验室 |
| 作者单位 | 1.Acad Sinica, Inst Oceanol, Qingdao 266071, Peoples R China 2.Harbin Inst Technol, Sch Ocean, Weihai 264209, Peoples R China |
| 推荐引用方式 GB/T 7714 | Hou, Yan-Hua,Wang, Quan-Fu,Ding, Ling,et al. attB site disruption in marine Actinomyces sp M048 via DNA transformation of a site-specific integration vector[J]. BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY,2008,50(Part 1):11-16. |
| APA | Hou, Yan-Hua,Wang, Quan-Fu,Ding, Ling,Li, Fu-Chao,&Qin, Song.(2008).attB site disruption in marine Actinomyces sp M048 via DNA transformation of a site-specific integration vector.BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY,50(Part 1),11-16. |
| MLA | Hou, Yan-Hua,et al."attB site disruption in marine Actinomyces sp M048 via DNA transformation of a site-specific integration vector".BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY 50.Part 1(2008):11-16. |
入库方式： OAI收割
来源：海洋研究所
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