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 0
In 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 CYP6N3v1 v3 (the full length cDNA sequences) and CYP6N3v4, CYP6N4v1 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 CYP6N3 and CYP6N4 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 CYP6N3 suggested that the multiplicity also exist in transcriptional initiation of CYP6 in mosquitoes. ⑥The diversity of CYP6 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.
出处
《中山医科大学学报》
CSCD
北大核心
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)
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
anopheles
aedes
culex
cytochrome P450
polymerase chain reaction methods
gene amplification
gene walking
gene expression regulation
transcription control
translation control
heterologous expression
