抗生链霉菌的黑色素操纵子：在革兰氏阴性菌中的表达及用作标记

The melanin operon of Streptomyces antibioticus: expression and use as a marker in gram-negative bacteria.

作者信息

Tseng H C, Lin C K, Hsu B J, Leu W M, Lee Y H, Chiou S J, Hu N T, Chen C W

机构信息

Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, Taiwan.

出版信息

Gene. 1990 Jan 31;86(1):123-8. doi: 10.1016/0378-1119(90)90124-a.

DOI:10.1016/0378-1119(90)90124-a

PMID:2107124

Abstract

The melC operon of Streptomyces antibioticus contains two genes, melC1 and melC2, necessary for the production of melanin pigment. We transferred the coding sequence of melC1 and melC2 to Escherichia coli plasmid pMTL23 such that its transcription was under the control of the lac promoter and melC1 was translationally fused to the lacZ alpha fragment. E. coli cultures containing this plasmid, pIF413, produced melanin after overnight incubation on 2YT agar supplemented with 0.1 mM CuCl2, 0.36 mM IPTG (or 0.2% lactose), and 2 mM tyrosine. Erwina carotovora could also be transformed by pIF413 to produce melanin. Two shuttle vectors were constructed: pLUS415 for E. coli and Streptomyces, and pLAF413 for E. coli and Xanthomonas campestris. These vectors confer melanin pigmentation in all the hosts that harbor them. The melC sequence provides the vectors with a convenient cloning marker for insertional or replacement inactivation.

摘要

抗生链霉菌的melC操纵子包含两个基因，即melC1和melC2，它们是黑色素产生所必需的。我们将melC1和melC2的编码序列转移至大肠杆菌质粒pMTL23，使其转录受lac启动子控制，且melC1与lacZα片段进行翻译融合。含有该质粒pIF413的大肠杆菌培养物在补充有0.1 mM氯化铜、0.36 mM异丙基-β-D-硫代半乳糖苷（或0.2%乳糖）和2 mM酪氨酸的2YT琼脂上过夜培养后产生黑色素。胡萝卜软腐欧文氏菌也可被pIF413转化以产生黑色素。构建了两种穿梭载体：用于大肠杆菌和链霉菌的pLUS415，以及用于大肠杆菌和野油菜黄单胞菌的pLAF413。这些载体在所有携带它们的宿主中都能产生黑色素沉着。melC序列为载体提供了一个方便的用于插入或替代失活的克隆标记。