| 委内瑞拉链霉菌中氯霉素生物合成基因的克隆 |
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| 引用本文： | 何建勇,姚新生,Leo.C.Vining. 委内瑞拉链霉菌中氯霉素生物合成基因的克隆[J]. 沈阳药科大学学报, 2006, 23(11): 731-734 |
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| 作者姓名： | 何建勇 姚新生 Leo.C.Vining |
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| 作者单位： | 1. 沈阳药科大学,制药工程学院,辽宁,沈阳,110016
2. Biology Department of Dalhousie University,B3H 4J1,Canada |
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| 摘 要： | 目的克隆委内瑞拉链霉菌(Streptomyces venezuelae)ISP5230中的氯霉素生物合成基因。方法以pabAB基因片段为探针,通过菌落杂交技术从该菌的基因文库中钓取相关基因。结果得到一7.5kbBamHIBamHI DNA片段,突变株的互补实验表明,该7.5kb DNA片段使氯霉素生物合成阻断变株CML-4恢复了氯霉素的生物合成。从野生菌株的染色体上删除该7.5 kb DNA片段中的一段3.5 kbNcoINcoI DNA片段后,野生菌株丧失了氯霉素的生物合成能力。结论该7.5 kb DNA片段含有氯霉素生物合成所需的基因。
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| 关 键 词： | 基因克隆 委内瑞拉链霉菌 氯霉素 |
| 文章编号： | 1006-2858(2006)11-0731-04 |
| 收稿时间： | 2006-02-08 |
| 修稿时间： | 2006-02-08 |
Cloning of the chloramphenicol biosynthesis genes of Streptomyces venezuelae-5230 |
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| HE Jian-yong,YAO Xin-sheng,Leo.C.Vining. Cloning of the chloramphenicol biosynthesis genes of Streptomyces venezuelae-5230[J]. Journal of Shenyang Pharmaceutical University, 2006, 23(11): 731-734 |
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| Authors: | HE Jian-yong YAO Xin-sheng Leo.C.Vining |
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| Affiliation: | 1. School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang 110016, China ; 2. Biology Department of Dalhousie University, B3H 4J1, Canada |
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| Abstract: | Objective To clone the chloramphenicol biosynthesis genes from Streptomyces venezuelae-5230.Methods The pabAB gene fragment of Streptomyces venezuelae5230 was used as a probe,searching for target genes from gene library of Streptomyces venezuelae-5230 by colony hybridization.Results A 7.5 kb BamHI-BamHI DNA fragment was obtained.Mutants complementation experiments showed that this(7.5 kb) DNA segment restored the chloramphenicol(Cm) production of a mutant cml-4.Deletion of the 3.5 kb NcoI-NcoI fragment in the 7.5 kb segment of the wild-type genomic DNA eliminated Cm production.Conclusions The 7.5kb BamHI-BamHI DNA fragment contains the genes necessary for the biosynthesis of chloramphenicol. |
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| Keywords: | gene cloning Streptomyces venezuelae chloramphenicol |
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