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链丝菌素生物合成基因簇的克隆及其异源表达

Cloning and heterologous expression of a streptothricin biosynthetic gene cluster
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摘要 通过基因组粘粒文库的高通量接合转移、异源表达和生物活性测定,结合DNA序列测定,从刺孢吸水链霉菌AA97026中筛选具有广谱抗菌活性的小分子化合物及其生物合成基因簇,获得1个对革兰氏阳性细菌和红酵母均有抑制活性的阳性克隆1H5,其部分DNA序列与链丝菌素生物合成基因相似。含1H5的异源链霉菌宿主的发酵液均能检测到链丝菌素的不同组份,表明1H5含有完整的链丝菌素生物合成基因簇。 To obtain small molecular antibacterial compound and its biosynthetic gene cluster,a ge- nomie cosmid library was screened by high-throughput conjugation, heterologous expression, bioassay, and combined with DNA sequencing. A positive clone 1145 expressing growth-inhibition activities against gram positive bacteria and Rhodotorufa was obtained from the genomic library of Streptomyces hygro- spinosus. The cosmid 1H5 harbored a cluster of genes highly similar to previously reported streptothricin biosynthetic gene clusters in terms of sequence and gene organization. Streptothricins were detected in the fermentation broth of Streptomyces expression hosts carrying 1H5, indicating that this clone con- tained an entire gene cluster for streptothricin biosynthesis. The cloning and heterologous expression of the biosynthetic gene cluster pave the way for further study of the biosynthesis, regulation and structural modification of streptothricin.
作者 陈萌 徐敏 王业民 白亭丽 毕德玺 邓子新 陶美凤
机构地区 上海交通大学生命科学技术学院/微生物代谢国家重点实验室 华中农业大学农业微生物学国家重点实验室
出处 《华中农业大学学报》 CAS CSCD 北大核心 2016年第2期41-47,共7页 Journal of Huazhong Agricultural University
基金 国家"973"计划项目(2009CB118905) 国家自然科学基金项目(FC31170084)
关键词 链霉菌 链丝菌素 基因组粘粒文库 高通量接合转移 异源表达 Streptomyces streptothricins genomic cosmid library high throughput conjugation heterologous expression
分类号 Q785 [生物学—分子生物学]
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参考文献20

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华中农业大学学报
2016年 第2期

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