Genetic transformation of marine actinomycete sp isolate m048 and expression of a recombinant plasmid carrying the apc gene
文献类型：期刊论文
| 作者 | Hou Yanhua; Li Fuchao; Qin Song; Wang Quanfu |
| 刊名 | Acta oceanologica sinica
 |
| 出版日期 | 2006 |
| 卷号 | 25期号:6页码:145-152 |
| 关键词 | Allophycocyanin Protoplast Intergeneric conjugantion Exconjugant Marine actinomycetes |
| ISSN号 | 0253-505X |
| 通讯作者 | Qin song(sqin@ms.qdio.ac.cn) |
| 英文摘要 | Optimal conditions for protoplasts formation of marine actinomycete sp. isolate m048 were described, dense and disperse mycelia were cultured in sggp medium, 0.5% glycine, lysozyme exposure (2 mg/cm(3), 37 degrees c, 40 min), and the concentration of sucrose in protoplast buffer was 0.4 mol/dm(3) for keeping the balance of osmotic pressure. using peg-mediated protoplasts transformation, the transformation frequency was 89 transformants per microgramme of pij702. meanwhile, an effective transformation procedure was established based on intergeneric conjugation from e. coli et12567 ( puz8002) using shuttle vectors ppm801, ppm803 and a phi c31-derived integration vector pij8600 containing orit and attp fragments. transformation frequencies were 5.30 x 10(-4) +/- 0.26 x 10(-4), 8.92 x 10(-4) +/- 0.19 x 10(-4) and 6.38 x 10(-5) +/- 0.41 x 10(-5), respectively. further, the heterologous expression of the allophycocyanin gene (apc) in the strain m048 was used to demonstrate this transformation system. sds - page and western blot analysis confirmed the expression of recombinant apc (rapc). |
| WOS关键词 | ESCHERICHIA-COLI ; INTERGENERIC CONJUGATION ; HIGH-FREQUENCY ; STREPTOMYCES ; DNA ; PURIFICATION ; ANTIBIOTICS ; DISCOVERY ; VECTORS ; CLONING |
| WOS研究方向 | Oceanography |
| WOS类目 | Oceanography |
| 语种 | 英语 |
| WOS记录号 | WOS:000243201300015 |
| 出版者 | CHINA OCEAN PRESS |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2378602 |
| 专题 | 中国科学院大学 |
| 通讯作者 | Qin Song |
| 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 |
| 推荐引用方式 GB/T 7714 | Hou Yanhua,Li Fuchao,Qin Song,et al. Genetic transformation of marine actinomycete sp isolate m048 and expression of a recombinant plasmid carrying the apc gene[J]. Acta oceanologica sinica,2006,25(6):145-152. |
| APA | Hou Yanhua,Li Fuchao,Qin Song,&Wang Quanfu.(2006).Genetic transformation of marine actinomycete sp isolate m048 and expression of a recombinant plasmid carrying the apc gene.Acta oceanologica sinica,25(6),145-152. |
| MLA | Hou Yanhua,et al."Genetic transformation of marine actinomycete sp isolate m048 and expression of a recombinant plasmid carrying the apc gene".Acta oceanologica sinica 25.6(2006):145-152. |
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来源：中国科学院大学
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