Cloning and characterization of the cytochrome P450 oxidoreductase gene from the zygomycete fungus Cunninghamella.
Cloning and characterization of the cytochrome P450 oxidoreductase gene from the zygomycete fungus Cunninghamella.
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接合菌 Cunninghamella 细胞色素 P450 氧化还原酶基因的克隆和表征。
DOI：
10.1006/bbrc.2000.2124
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Loper,JC
中科院分区：
文献类型：
--
作者：
Yadav,JS;Loper,JC
丝状真菌Cunninghamella利用细胞色素P450系统代谢多种多芳族和脂肪族污染物以及多种药物，但先前分离该真菌P450系统成分的尝试通常不成功。我们报道了两个广泛研究的物种，秀丽隐杆线虫和棘球线虫的细胞色素P450氧化还原酶（CPR）基因。以已知PCR扩增子为探针，用退化引物筛选基因组文库，获得秀丽隐杆线虫的CPR基因。2420bp编码区包含两个内含子（149 bp和138 bp）。Northern blot分析显示，CPR基因在秀丽隐杆线虫中转录表达，似乎可以被烷烃底物正十四烷诱导。对推导出的秀丽隐杆线虫CPR （710 aa）的系统发育比较表明，它与动物CPR（41-42%）的亲缘关系比与酵母（38-41%）和植物（35-36%）的亲缘关系更密切。从棘球蚴中分离到一个含有大部分CPR基因同源物的2074 bp序列。
The filamentous fungus Cunninghamella utilizes cytochrome P450 system(s) in the metabolism of a broad range of polyaromatic and aliphatic pollutants and a variety of drugs, but prior attempts at isolation of P450 system components of this fungus have been generally unsuccessful. We report upon the cytochrome P450 oxidoreductase (CPR) gene from two widely studied species, C. elegans and C. echinulata. The C. elegans CPR gene was obtained by screening a genomic library using as probe a PCR amplicon obtained with degenerate primers based on known CPRs. The 2420 bp coding region contained two apparent introns (149 bp and 138 bp). Northern blot analysis showed that the CPR gene is transcriptionally expressed in C. elegans and appears to be inducible by an alkane substrate, n-tetradecane. Phylogenetic comparison of the deduced C. elegans CPR (710 aa) suggested that it is more closely related to animal CPRs (41–42%) than to yeast (38–41%) and plant (35–36%) forms. A 2074 bp sequence containing most of the CPR gene homolog from C. echinulata was also isolated.