THE CDNA AND DEDUCED PROTEIN-SEQUENCE OF HOUSE-FLY NADPH-CYTOCHROME P450 REDUCTASE
THE CDNA AND DEDUCED PROTEIN-SEQUENCE OF HOUSE-FLY NADPH-CYTOCHROME P450 REDUCTASE
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DOI：
10.1016/0965-1748(93)90051-s
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发表时间：
1993-06-01
影响因子：
3.8
通讯作者：
FEYEREISEN, R
中科院分区：
文献类型：
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作者：
KOENER, JF;CARINO, FA;FEYEREISEN, R
纯化的家蝇NADPH-细胞色素P450还原酶的抗血清被用来选择从表达文库的苯巴比妥处理的家蝇的cDNA克隆。获得了一个1841 bp的cDNA片段，其中含有一个TAG终止密码子、一个共有的多聚腺苷酸化位点和269 bp的3'端非翻译序列。基因组克隆的测序与mRNA测序偶联产生包括起始ATG的完整编码序列。得到的2013个核苷酸的开放阅读框编码671个残基的蛋白质。天然还原酶脱辅基蛋白的分子量为76，366，全酶（即含有1 mol FAD和FMN）的推导分子量为77，608。家蝇P450还原酶蛋白的序列与兔肝的高度相似，总体氨基酸位置一致性为54.5%，真核生物P450还原酶之间的总体一致性约为25%。P450还原酶基因位于第三染色体上，其长度为19-23 kb。
Antisera to purified house fly NADPH-cytochrome P450 reductase were used to select cDNA clones from an expression library of abdomens of phenobarbital-treated house flies. A partial cDNA of 1841 bp containing a TAG termination codon, a consensus polyadenylation site and 269 bp of 3' untranslated sequence was obtained. Sequencing of a genomic clone coupled with mRNA sequencing yielded the complete coding sequence including the starting ATG. The resulting open reading frame of 2013 nucleotides codes for a protein of 671 residues. The native reductase apoprotein has a molecular weight of 76,366 and the deduced molecular weight of the holoenzyme (i.e. with 1 mol of FAD and FMN) is 77,608. The sequence of the house fly P450 reductase protein is highly similar to that of rabbit liver, the overall amino acid positional identity is 54.5% and the overall identity among eukaryotic P450 reductases is about 25%. The P450 reductase gene of 19-23 kb was located on chromosome III, as shown by comparison of RFLP-patterns of the P450 reductase gene in two house fly strains and their hybrids.