CLONING AND EXPRESSION OF DAUNORUBICIN BIOSYNTHESIS GENES FROM STREPTOMYCES-PEUCETIUS AND STREPTOMYCES-PEUCETIUS SUBSP CAESIUS
CLONING AND EXPRESSION OF DAUNORUBICIN BIOSYNTHESIS GENES FROM STREPTOMYCES-PEUCETIUS AND STREPTOMYCES-PEUCETIUS SUBSP CAESIUS
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DOI：
10.1128/jb.172.6.3427-3434.1990
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发表时间：
1990-06-01
影响因子：
3.2
通讯作者：
HUTCHINSON, CR
中科院分区：
文献类型：
--
作者：
OTTEN, SL;STUTZMANENGWALL, KJ;HUTCHINSON, CR
从柔红霉素产生源链霉菌peucetius和S. peucetius亚种中克隆了重要的抗肿瘤药物柔红霉素（daunomycin）和阿霉素（阿霉素）的生物合成基因。利用actii /tcmIa和actIII聚酮合成酶基因探针对多柔比星生产商caesius进行检测。在cosmid载体上克隆的DNA进行限制性定位和Southern分析，确定了该DNA代表了S. peucetius亚种的三个不重叠区域。caesius基因组。这三个区域加上另外一个与相同探针杂交的区域存在于S. peucetius基因组中，如先前报道（K.J. Stutzman-Engwall和c.r. Hutchinson, Proc. Natl）。学会科学。《美国科学》86:3135-3139,1989)。S. lividans、S. peucetius ATCC 29050、S. peucetius subsp中部分区域代表性克隆的功能分析。该菌株ATCC 27952及其两个被阻断的突变体（菌株H6101和H6125）表明，许多抗生素生产基因存在于IV组克隆所代表的DNA区域。这个结论是基于。epsilon的产生。-rhodomycinone是柔红霉素途径的关键中间体，在某些S. lividans转化体中，以及在菌株H6101和H6125中阻断柔红霉素生物合成的突变的明显互补。菌株29050、27952和H6125的一些转化体表现出大量的。epsilon过剩。红红霉素和柔红霉素。
Genes for the biosynthesis of daunorubicin (daunomycin) and doxorubicin (adriamycin), important antitumor drugs, were cloned from Streptomyces peucetius (the daunorubicin producer) and S. peucetius subsp. caesius (the doxorubicin producer) by use of the actI/tcmIa and actIII polyketide synthease gene probes. Restriction mapping and Southern analysis of the DNA cloned in a cosmid vector established that the DNA represented three nonoverlapping regions of the S. peucetius subsp. caesius genome. These three regions plus an additional one that hybridized to the same probes are present in the S. peucetius genome, as reported previously (K.J. Stutzman-Engwall and C. R. Hutchinson, Proc. Natl. Acad. Sci. USA 86:3135-3139, 1989). Functional analysis of representative clones from some of these regions in S. lividans, S. peucetius ATCC 29050, S. peucetius subsp. caesius ATCC 27952, and two of its blocked mutants (strains H6101 and H6125) showed that many of the antibiotic production genes reside in the region of DNA represented by the group IV clones. This conclusion is based on the production of .epsilon.-rhodomycinone, a key intermediate of the daunorubicin pathway, in certain S. lividans transformants and on the apparent complementation of mutations that block daunorubicin biosynthesis in strains H6101 and H6125. Some of the transformants of strains 29050, 27952, and H6125 exhibited substantial overproduction of .epsilon.-rhodomycinone and daunorubicin.