本文選題：白紋伊蚊 + 吸血行為��； 參考：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2010年碩士論文

【摘要】： 白紋伊蚊Aedes albopictus是我國登革熱和登革出血熱的主要傳播媒介,也是城市滅蚊的主要對象。目前,對白紋伊蚊進行化學(xué)防治一直是控制登革熱流行的主要手段,然而,隨著蚊蟲抗藥性的出現(xiàn)和抗藥性水平的不斷增高,防治效果受到影響,同時,化學(xué)殺蟲劑對環(huán)境造成的污染也一直是無法避免的問題。因此,近年來,如何有效利用白紋伊蚊的生理行為特點,發(fā)展新的防治方法已日益受到關(guān)注。 以往的研究發(fā)現(xiàn),蚊蟲對宿主的吸血習(xí)性是以嗅覺為基礎(chǔ),并由遺傳基因所決定。蚊蟲吸血行為的嗅覺機制中,多種蛋白參與了蚊蟲對宿主氣味分子的識別和定位過程。在這些蛋白中,氣味結(jié)合蛋白(odorant-binding protein,OBP)數(shù)量最多,它位于觸角感器的淋巴液中,直接與外界環(huán)境中成千上萬的氣味分子接觸。氣味分子一般為脂溶性的化合物,必須與OBP結(jié)合才能穿過親水性的觸角感器淋巴液,到達神經(jīng)樹突膜上的氣味受體,產(chǎn)生動作電位。OBP識別和結(jié)合氣味分子是蚊蟲感受宿主氣味的第一步,在蚊蟲尋找血源定位宿主的過程中起重要作用。目前國內(nèi)外對蚊蟲OBP的研究主要集中在岡比亞按蚊上,也有少量關(guān)于埃及伊蚊、致倦庫蚊和白紋伊蚊的報道,其中白紋伊蚊OBP的研究報道僅有3個基因序列被識別且沒有更深入的研究。因此,白紋伊蚊的高度嗜人血習(xí)性可能與哪些OBP基因有關(guān),又是哪些OBP基因參與了白紋伊蚊重復(fù)吸血的獨特生理過程,這些都不清楚。而OBP基因作用于白紋伊蚊嗜血行為中感受宿主氣味的第一步,因此找到這些與其吸血行為相關(guān)的OBP基因并明確其功能是揭示白紋伊蚊吸血行為嗅覺機制的突破口,對于闡明白紋伊蚊吸血習(xí)性的內(nèi)在分子機制具有重要的意義。更重要的是,對白紋伊蚊吸血習(xí)性內(nèi)在機制的闡明,也為發(fā)展新的防治方法干擾白紋伊蚊-人宿主之間的聯(lián)系,從而阻斷蚊媒病的流行提供了新的思路和途徑。 本研究結(jié)果如下: (1)通過構(gòu)建白紋伊蚊雌蚊的觸角cDNA文庫,克隆得到2個氣味結(jié)合蛋白核酸序列,分別與埃及伊蚊OBP 56a和56e具有大于80%的相似度,命名為AealOBP1和AealOBP2;另外還得到2個嗅覺受體、1個氣味受體、1個味覺受體和3個荷爾蒙受體的基因核酸序列,都與埃及伊蚊相關(guān)基因的相似性最高。 (2)通過設(shè)計簡并引物,對白紋伊蚊雌蚊觸角cDNA進行擴增,得到6個OBP基因序列,其中1個與文庫中發(fā)現(xiàn)的AealOBP1序列相重合,剩余5個OBP基因序列分別與埃及伊蚊、致倦庫蚊和岡比亞按蚊OBP基因序列具有大于75%的相似性,命名為AealOBP3至AealOBP7; (3)實時定量PCR檢測10個OBP(AealOBP1、AealOBP3～7、AealOBP46、AealOBP67和AealOBP68)基因在雌蚊吸血前后和雄蚊頭部起始表達量的差異,其中AealOBP1、AealOBP4和AealOBP5基因在吸血前后雌蚊頭部和雌雄蚊頭部表達無顯著差異;飽血雌蚊頭部AealOBP6、AealOBP46和AealOBP68的表達量明顯多于未吸血雌蚊頭部,且AealOBP67和AealOBP46在雄蚊頭部的表達量也明顯多于未吸血雌蚊;AealOBP3和AealOBP7在吸血后雌蚊頭部表達量顯著降低,而AealOBP7在未吸血雌蚊頭部的表達量又明顯多于雄蚊。
[Abstract]:Aedes albopictus Aedes albopictus is the main medium of dengue and dengue hemorrhagic fever in China, and it is also the main object of urban mosquito control. At present, chemical control of Aedes albopictus has been the main means to control the epidemic of dengue fever. However, with the emergence of mosquito resistance and the increasing resistance level, the control effect has been affected. At the same time, the environmental pollution caused by chemical pesticides has also been an unavoidable problem. Therefore, in recent years, how to effectively use the physiological behavior of Aedes albopictus and the development of new control methods have been paid more and more attention.
Previous studies have found that the blood sucking habit of mosquitoes to the host is based on the olfactory sense and is determined by genetic genes. In the olfactory mechanism of the mosquito sucking behavior, a variety of proteins participate in the process of identification and localization of mosquitoes to host odorants. In these proteins, the number of odorant-binding protein, OBP is the most, and its number is the most. In the lymph fluid of the antennae, it is directly contacted with thousands of odorant molecules in the external environment. Odorant molecules are generally fat soluble compounds, which must be combined with OBP to pass through the hydrophilic tentacle lymph, to reach the odorant receptors on the nerve dendrite membrane, and to produce dynamic potential.OBP recognition and combination of odorant molecules that are mosquitoes. The first step of host odor plays an important role in the process of locating the host by the mosquito in search of the source of the source of the blood. At present, the study of mosquitoes at home and abroad is mainly concentrated in Anopheles Gambia, and a small amount of reports on Aedes aegypti, Culex pipiens and Aedes albopictus, of which only 3 gene sequences have been identified and only identified in the study of Aedes albopictus OBP. There is no more in-depth study. Therefore, the high eosinophilic habit of Aedes albopictus may be related to which OBP genes and which OBP genes participate in the unique physiological process of the Aedes albopictus in repeated bloodsucking, which is not clear. And the OBP gene acts on the first step of the Aedes albopictus to feel the host odor in the bloodthirsty behavior of Aedes albopictus. The OBP gene related to the blood sucking behavior and its function are a breakthrough in revealing the olfactory mechanism of Aedes albopictus, which is important to elucidate the intrinsic molecular mechanism of the blood sucking habit of Aedes albopictus, and more importantly, the internal mechanism of the blood sucking habit of Aedes albopictus, and the development of new control methods to interfere with the Aedes albopictus. The link between human hosts, thereby blocking the prevalence of mosquito borne diseases, has provided new ideas and approaches.
The results of this study are as follows:
(1) by constructing the antennae cDNA Library of Aedes albopictus, 2 odorant binding protein nucleic acid sequences were cloned, and the similarity between the OBP 56a and 56e of Aedes aegypti was greater than 80%, respectively, named AealOBP1 and AealOBP2, and 2 olfactory receptors, 1 odorant receptors, 1 taste receptors and 3 hormone receptor sequences were also obtained. All of them have the highest similarity to Aedes aegypti related genes.
(2) through the design of degenerate primers, 6 OBP gene sequences were amplified for the cDNA of Aedes albopictus, and 1 of them coincided with the AealOBP1 sequences found in the library. The remaining 5 OBP sequences were similar to the OBP gene sequences of Aedes aegypti, Culex pipiens tirenopus and Anopheles Gambia, respectively, named AealOBP3 to AealOB P7;
(3) the difference in the initial expression of 10 OBP (AealOBP1, AealOBP3 to 7, AealOBP46, AealOBP67 and AealOBP68) genes in the head of the male and male mosquitoes was detected by real time quantitative PCR. There was no significant difference in the expression of AealOBP1, AealOBP4 and AealOBP5 genes between the head of the female and male and male mosquitoes before and after the blood sucking; the head of the female mosquito satiety was AealOBP6, AealOBP46. The expression of and AealOBP68 was significantly more than the head of the non blood sucking female mosquitoes, and the expression of AealOBP67 and AealOBP46 in the head of the male mosquitoes was significantly more than that of the non blood sucking females. The expression of AealOBP3 and AealOBP7 in the head of the female mosquitoes decreased significantly after the blood sucking, and the expression of AealOBP7 in the head of the non blood sucking female mosquitoes was significantly more than that of the male mosquito.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R384.1

【引證文獻】

相關(guān)期刊論文 前2條

1 李躍;劉乾;鄭學(xué)禮;;白紋伊蚊氣味結(jié)合蛋白的研究進展[J];熱帶醫(yī)學(xué)雜志;2011年11期

2 趙大川;鄭學(xué)禮;;白紋伊蚊氣味結(jié)合蛋白基因OBP68的克隆鑒定及其在蚊不同發(fā)育時期表達分析[J];中國病原生物學(xué)雜志;2013年03期

，

本文編號：1921299