CLONING OF THE ASPERGILLUS-PARASITICUS APA-2 GENE ASSOCIATED WITH THE REGULATION OF AFLATOXIN BIOSYNTHESIS
CLONING OF THE ASPERGILLUS-PARASITICUS APA-2 GENE ASSOCIATED WITH THE REGULATION OF AFLATOXIN BIOSYNTHESIS
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DOI：
10.1128/aem.59.10.3273-3279.1993
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发表时间：
1993-10-01
影响因子：
4.4
通讯作者：
PAYNE, GA
中科院分区：
文献类型：
--
作者：
CHANG, PK;CARY, JW;PAYNE, GA
一个被命名为apa - 2的寄生曲霉基因被鉴定为与黄曲霉毒素生物合成相关的调控基因。在含有黄曲霉毒素生物合成基因nor - 1和ver - 1的粘粒DNA转化真菌菌株后，基于途径中间体的过量产生，克隆了apa - 2基因。用含有apa - 2的一个5.5 - kb HindIII - XbaI DNA片段转化一株积累O - 甲基柄曲霉素的菌株——寄生曲霉SRRC 2043，导致所分析的所有黄曲霉毒素途径中间体过量产生。与去甲氧基胶霉毒素和柄曲霉素转化相关的特定酶活性大约增加了两倍。发现apa - 2基因能使黄曲霉afl - 2突变株恢复黄曲霉毒素的产生，这表明apa - 2与afl - 2在功能上是同源的。寄生曲霉apa - 2基因的DNA序列与黄曲霉afl - 2基因（G. A. 佩恩，G. J. 奈斯特龙，D. 巴特纳格尔，T. E. 克利夫兰和C. P. 沃洛舒克，《应用与环境微生物学》59:156 - 162, 1993）的比较表明，它们具有>95%的DNA同源性。粘粒亚克隆的物理图谱显示apa - 2距离ver - 1约8 kb。
An Aspergillus parasiticus gene, designated apa-2, was identified as a regulatory gene associate with aflatoxin biosynthesis. The apa-2 gene was cloned on the basis of overproduction of pathway intermediates following transformation of fungal strains with cosmid DNA containing the aflatoxin biosynthetic genes nor-1 and ver-1. Transformation of an 0-methylsterigmatocystin-accumulating strain, A. parasiticus SRRC 2043, with a 5.5-kb HindIII-XbaI DNA fragment containing apa-2 resulted in overproduction of all aflatoxin pathway intermediates analyzed. Specific enzyme activities associated with the conversion of norsolorinic acid and sterigmatocystin were increased approximately twofold. The apa-2 gene was found to complement an A. flavus afl-2 mutant strain for aflatoxin production, suggesting that apa-2 is functionally homologous to afl-2. Comparison of the A. parasiticus apa-2 gene DNA sequence with that of the A. flavus afl-2 gene (G. A. Payne, G. J. Nystorm, D. Bhatnagar, T. E. Cleveland, and C. P. Woloshuk, Appl. Environ. Microbiol. 59:156-162, 1993) showed that they shared >95% DNA homology. Physical mapping of cosmid subclones placed apa-2 approximately 8 kb from ver-1.