中华按蚊CYP6基因家族的鉴定、特征及与抗药性相关的表达分析

发布时间：2018-05-26 09:50

本文选题：中华按蚊 + CYP6　；参考：《重庆师范大学》2014年硕士论文

【摘要】：蚊媒传染病严重威胁公众健康，其中疟疾危害最为严重。仅2012年，全球就有2.07亿人感染疟疾，，造成约62.7万人死亡。喷洒杀虫剂以及悬挂经杀虫剂浸泡的蚊帐为防控疟疾最常用手段。自上世纪50年代开始，各类杀虫剂被用于媒介蚊虫的控制，然而随着杀虫剂的广泛使用，在全球多达64个国家已发现蚊虫对杀虫剂产生抗药性。蚊虫对杀虫剂产生抗药性的主要原因是解毒酶的过量表达。P450是蚊虫体内3大类解毒酶之一，CYP6家族基因与杀虫剂抗药性密切相关。鉴定蚊虫抗药性相关基因，研究其功能，阐释蚊虫抗药性分子机理对于蚊媒疾病的有效控制具有指导意义。本研究利用HMM和双向Blast的方法鉴定了中华按蚊CYP6家族基因，对其基因结构和特征进行了生物信息学分析，并对安徽、云南和重庆的对溴氰菊酯敏感和产生抗药性的中华按蚊构建的表达谱进行了初步分析，得到的主要结论如下： ①在中华按蚊中共鉴定出22个CYP6家族基因。中华按蚊基因组中鉴定出的22个CYP6家族基因分属于CYP6AA、CYP6AD、CYP6AF、CYP6AG、CYP6AH、CYP6M、CYP6N、CYP6P、CYP6R、CYP6Y和CYP6Z这11个亚家族。 ②中华按蚊CYP6家族基因与冈比亚按蚊CYP6家族基因内含子数目和内含子型相同。除CYP6AG和CYP6AH亚家族之外的9个亚家族仅含有1个相位1型内含子；而CYP6AG亚家族含有3个内含子，内含子1为相位2型内含子，内含子2为相位0型内含子，内含子3为相位1型内含子，CYP6AH亚家族则含有4个内含子，分别为相位0型、相位2型、相位0型和相位1型内含子。 ③中华按蚊基因组CYP6家族有2个基因簇。中华按蚊基因组数据分析显示CYP6Y亚家族基因与AsCYP6N3、AsCYP6R1构成了1个基因簇；中华按蚊中AsCYP6AD1与AsCYP6P4、AsCYP6P1和AsCYP6P2构成了另1个簇。 ④中华按蚊表达谱中CYP6AA1、CYP6AA2、CYP6M2、CYP6P3、CYP6P5和CYP6Z1有差异表达。CYP6P3和CYP6P5在安徽的抗药性雌蚊中上调表达，CYP6AA1、CYP6AA2、CYP6P3和CYP6P5在云南的抗药性雌蚊中上调表达，CYP6M2和CYP6Z1在重庆的抗药性雌蚊中下调表达。
[Abstract]:Mosquito-borne infectious diseases pose a serious threat to public health, of which malaria is the most serious. In 2012 alone, 207 million people worldwide were infected with malaria, killing about 627000 people. Spraying insecticides and hanging insecticide-soaked mosquito nets are the most common means of malaria control. Since the 1950s, various insecticides have been used for vector mosquito control. However, with the widespread use of insecticides, mosquitoes have been found to be resistant to insecticides in up to 64 countries around the world. The main reason for mosquito resistance to insecticides is that the over-expression of detoxifying enzymes. P450 is one of the three major detoxifying enzymes in mosquitoes. CYP6 family genes are closely related to insecticide resistance. The identification of mosquito resistance related genes, the study of their function, and the elucidation of the molecular mechanism of mosquito resistance have guiding significance for the effective control of mosquito-borne diseases. In this study, the genes of CYP6 family of Anopheles sinensis were identified by HMM and two-way Blast, and their gene structure and characteristics were analyzed by bioinformatics. The expression profiles of Anopheles sinensis which were sensitive to deltamethrin and resistant to drug were preliminarily analyzed in Yunnan and Chongqing. The main conclusions were as follows: 1 A total of 22 CYP6 family genes were identified in Anopheles sinensis. The 22 CYP6 family genes identified from the genome of Anopheles sinensis belong to 11 subfamilies, CYP6AAAA-CYP6ADAG-CYP6AAGG, CYP6AHG, CYP6AHN, CYP6P6N, CYP6RGY, and CYP6Z, which belong to the 11 subfamilies of CYP6AA-Anopheles sinensis (Anopheles sinensis). 2Anopheles sinensis CYP6 family gene was the same as Anopheles gambiae CYP6 gene intron number and intron type. Except for the CYP6AG and CYP6AH subfamilies, the 9 subfamilies contained only one phase 1 intron, while the CYP6AG subfamily contained 3 introns, in which intron 1 was phase 2 intron, and intron 2 was phase 0 intron. The CYP6AH subfamily of intron 3 has four introns: phase 0, phase 2, phase 0 and phase 1. 3Genomic CYP6 family of Anopheles sinensis has two gene clusters. The genomic data analysis of Anopheles sinensis showed that the subfamily genes of CYP6Y and AsCYP6N3ACYP6R1 constituted a gene cluster, while AsCYP6AD1 and AsCYP6P4Anopheles sinensis formed another cluster. (4) CYP6AA1, CYP6AA2Anopheles sinensis and CYP6M3CYP6P5 were differentially expressed with CYP6Z1. CYP6P3 and CYP6P5 up-regulated the expression of CYP6A1CY CYP6AA2CYP6P3 and CYP6P6P6P3 and CYP6P5 down-regulated the expression of CYP6M2 and CYP6Z1 in resistant females in Chongqing.
【学位授予单位】：重庆师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R384.1

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