寄生曲霉中与黄曲霉毒素生物合成调控相关的apa-2基因的克隆
Cloning of the Aspergillus parasiticus apa-2 gene associated with the regulation of aflatoxin biosynthesis.
作者信息
Chang P K, Cary J W, Bhatnagar D, Cleveland T E, Bennett J W, Linz J E, Woloshuk C P, Payne G A
机构信息
Department of Cell and Molecular Biology, Tulane University, New Orleans, Louisiana 70118.
出版信息
Appl Environ Microbiol. 1993 Oct;59(10):3273-9. doi: 10.1128/aem.59.10.3273-3279.1993.
An Aspergillus parasiticus gene, designated apa-2, was identified as a regulatory gene associated 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 O-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.
一种寄生曲霉基因，命名为apa - 2，被鉴定为与黄曲霉毒素生物合成相关的调控基因。基于用含有黄曲霉毒素生物合成基因nor - 1和ver - 1的黏粒DNA转化真菌菌株后途径中间体的过量产生，克隆了apa - 2基因。用含有apa - 2的5.5 kb HindIII - XbaI DNA片段转化积累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。与诺索尔菌素酸和柄曲霉素转化相关的特定酶活性增加了约两倍。发现apa - 2基因可弥补黄曲霉afl - 2突变菌株的黄曲霉毒素产生，这表明apa - 2在功能上与afl - 2同源。将寄生曲霉apa - 2基因的DNA序列与黄曲霉afl - 2基因的DNA序列（G. A. Payne、G. J. Nystorm、D. Bhatnagar、T. E. Cleveland和C. P. Woloshuk，《应用与环境微生物学》59:156 - 162，1993）进行比较，结果表明它们的DNA同源性大于95%。黏粒亚克隆的物理图谱显示apa - 2距离ver - 1约8 kb。
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