CDNA AND DEDUCTED PROTEIN-SEQUENCE OF CYP6D1 - THE PUTATIVE GENE FOR A CYTOCHROME-P450 RESPONSIBLE FOR PYRETHROID RESISTANCE IN-HOUSE FLY
CDNA AND DEDUCTED PROTEIN-SEQUENCE OF CYP6D1 - THE PUTATIVE GENE FOR A CYTOCHROME-P450 RESPONSIBLE FOR PYRETHROID RESISTANCE IN-HOUSE FLY
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DOI：
10.1016/0965-1748(94)00066-q
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发表时间：
1995-02-01
影响因子：
3.8
通讯作者：
SCOTT, JG
中科院分区：
文献类型：
--
作者：
TOMITA, T;SCOTT, JG
家蝇微粒体细胞色素P450被称为P450(LPR)，参与P450单加氧酶介导的拟除虫菊酯抗性，在抗药性LPR品系中的表达水平是敏感品系的8倍。根据已知的P450(LPR)多肽序列，用简并引物进行聚合酶链式反应(PCR)扩增，扩增出内含子序列，用单侧特异性聚合酶链式反应(PCR)扩增出其余序列。从3个重叠的聚合酶链式反应产物中扩增出1.8kb的cDNA序列，编码516个残基的P450蛋白(M(R)59,182)。该基因被命名为P450基因超家族中的CYP6D1。该基因与蝴蝶、家蝇和果蝇的同源性最高(28.2-29.8%)，与大鼠的同源性为24.7%(P<0.05)。推导出的蛋白质序列包含一个疏水的N-末端区域和保守的序列，被认为参与了血红素结合和电子供体-蛋白质相互作用。将其与四个最相似的蛋白质(CYP6B1、CYP6A1、CYP6A2和CYP3A1)进行比较，发现在两个可能的底物结合区的序列中存在大量的残基。基因组内含子74、66和的3个内含子，分别具有5‘-GT和AG-3’端，分裂了基因组中的细胞色素P6D1编码区。结果表明，该基因可能是P450(LPR)基因。
A microsomal cytochrome P450 from the house fly (Musca domestica), termed P450(lpr), is involved in P450 monooxygenase-mediated pyrethroid resistance and is expressed at 8-fold higher levels in the insecticide resistant LPR strain compared to a susceptible strain. An internal cDNA sequence was amplified by polymerase chain reaction (PCR) using degenerate primers based on known P450(lpr) polypeptide sequences, and the remainder of the sequence was amplified by single side-specific PCR. A 1.8 kb cDNA sequence was obtained from 3 overlapping PCR products, with an open reading frame encoding a P450 protein of 516 residues (M(r) 59,182). This gene has been designated CYP6D1 within the P450 gene superfamily. CYP6D1 exhibits most similarity (28.2-29.8% positional identity) to butterfly CYP6B1, house fly CYP6A1 and Drosophila CYP6A2, and also exhibits comparable similarity (24.7% identity) to rat CYP3A1. The deduced protein sequence contains a hydrophobic N-terminal region and conserved sequences thought to be involved in heme-binding and electron donor-protein interactions. Comparison of CYP6D1 with its four most similar proteins (CYP6B1, CYP6A1, CYP6A2 and CYP3A1) reveals the presence of extensive stretches of residues in an alignment row in 2 possible substrate-binding regions. Three introns of 74, 66 and 64 bp, having 5'-GT and AG-3' ends, split the CYP6D1 coding region in genomic DNA. Results indicate that CYP6D1 is likely the P450(lpr) gene.