This work was supported by The National Natural Science Fundation of China (39970666).
采用一对昆虫细胞色素P450简并引物,以反转录-聚合酶链反应从淡色库蚊对溴氰菊酯敏感品系和抗性品系成虫RNA扩增到约485 bp和510 bp两个片段,将这两个片段与PinPointTMXa-1 T质粒重组,然后克隆至大肠杆菌JM109菌株,筛选获得68个阳性克隆;其中24个阳性克隆测序后与GenBank资料对照,显示为细胞色素P450新序列;分子系统学研究显示,24个新基因(等位基因)分别属CYP4家族CYP4C、CYP4D、CYP4H和CYP4J等4个亚家族,其已由细胞色素P450命名委员会命名和GenBank登录上网;其中CYP4C23可能是一个假基因,CYP4H13具有一段58个碱基长度的内含子,CYP4J4V1在近3′端具有一个终止密码子TAG.
24 new cDNA sequences encoding cytochrome P450 were amplified respectively from deltamethrin-susceptible and -resistant strains of the mosquito, Culex pipiens pallens, with a pair of degenerate primers according to the conservative amino acid sequences of CYP4 in insects by RT-PCR. Studies of molecular systematics show that the 24 new genes (alleles) belong to CYP4C,CYP4D,CYP4H and CYP4J subfamilies of the CYP4 family, and they were named by Cytochrome P450 Nomenclature Committee. Among the new genes(alleles),CYP4C23 may be a pseudogene, CYP4H13 has a retained intron 58 nucleotides in length, and CYP4J4V1 has a stop coden(TAG) in frame near the 3′-end.
朱昌亮,李建民,高晓红,田海生,李秀兰,沈波,吴观陵.淡色库蚊细胞色素P450基因研究[J].生物化学与生物物理进展,2001,28(1):61-66
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