蚊虫细胞色素P450基因系列研究  

A Series of Studies on Cytochrome P450 Genes in Mosquitoes

作　　者：黄炯烈[1] 周国理[1] 吴瑜[1] 葛春喜[1] 王玲[1] 

机构地区：[1]中山医科大学寄生虫学教研室,广东广州510080

出　　处：《中山医科大学学报》2001年第6期401-405,共5页Academic Journal of Sun Yat-sen University of Medical Sciences

基　　金：广东省自然科学基金资助项目 ( 970 0 92 );"2 11工程"重点学科建设基金资助项目 ( 9812 4)

摘　　要：报告中山医科大学寄生虫学教研室医学昆虫学室近 5年来有关蚊虫细胞色素P45 0基因 (CYP45 0 )系列研究结果 ,包括 :①通过简并引物PCR分别从致倦库蚊、白纹伊蚊及中华按蚊中获得 3个、2个、2个CYP4家族成员基因片段 ;通过简并引物PCR及RT PCR从白纹伊蚊中获得 16个CYP6家族成员基因片段 ;②利用cDNA末端快速扩增 (RACE)法获得了白纹伊蚊CYP6家族 3个新的全长cDNA序列及 7个超过其全长一半以上的cDNA序列。GenBankaccessionnumber分别为AF2 83836 AF2 83838(全长cDNA序列 )和AF2 84782 AF2 84783(非全长cDNA序列 ) ,被国际P45 0命名委员会正式命名为CYP 6N3v1 v3(全长cDNA序列 )和CYP 6N3v4、CYP 6N4v1 v6 ,并证实这些新基因同时存在于白纹伊蚊溴氰菊酯抗性株及敏感株中 ;③运用基因步移方法从白纹伊蚊溴氰菊酯抗性株中扩增出CYP 6N3上游 3 0 76bp调控序列 ;④对白纹伊蚊CYP 6N3、CYP 6N4非翻译区序列功能分析显示 :昆虫CYP45 0与其它真核mRNA翻译起始及终止机制相似 ,但具有多态性的特点 ;⑤对白纹伊蚊CYP 6N3上游启动子区序列特征与功能分析显示 :蚊虫中CYP45 0转录起始存在着复杂性 ;⑥证实蚊虫中 CYP6存在多样性 ,并分析讨论了此种多样性形成的可能机制 ;⑦分析讨论了蚊虫细胞色素P45 0In this report, a series of research results of mosquito cytochrome P450 genes (CYP450) during the last 5 years were reviewed by the research group of medical arthropods in Department of Parasitology, Sun Yat sen University of Medical Sciences. The content included: ①7 CYP4 gene (CYP4) fragments were cloned from mosquitoes Culex quinquefasciatus, Aedes albopictus (Ae.albopictus) and Anopheles sinensis by degenerated PCR technique, respectively; 16 CYP6 gene (CYP6) fragments were isolated from mosquitoes Ae.albopictus by degenerate PCR and degenerate RT PCR techniques. ②3 new full length cDNAs and 7 new cDNA fragments (each more than half of the full length cDNA) were obtained from Ae.albopictus by RACE method, which are submitted by GenBank Database (accession number: AF283836 AF283838 and AF284782 AF284783), and named as CYP6N3v1 v3 (the full length cDNA sequences) and CYP6N3v4, CYP6N4v1 v6 (the non full length cDNA sequences) by International P450 Nomenclature Committee, respectively. ③A 3 076 bp 5′ flanking nucleotide sequence were amplified from deltamethrin resistant Ae.albopictus by genomic walking method. ④The obtained 5′ UTR and 3′ UTR of CYP6N3 and CYP6N4 were analyzed. The results showed that the translation start and stop mechanisms of CYP450 in insects are similar to that of other eukaryo tic mRNAs, but characterized by multiplicity. ⑤The analysis on the obtained promoter sequence of CYP6N3 suggested that the multiplicity also exist in transcriptional initiation of CYP6 in mosquitoes. ⑥The diversity of CYP6 in mosquitoes was verified and its possible causes were discussed. ⑦The molecular evolutionary mechanism of CYP450 in mosquitoes was discussed. ⑧Our further research goals were introduced.

关 键 词：按蚊属 伊蚊属 库蚊属 细胞色素P450 聚合酶链反应 蚊虫 基因扩增 基因步移 基因表达调控 转录调控 

分 类 号：R384.1[医药卫生—医学寄生虫学]