【摘要】：瘧疾是嚴(yán)重危害人體健康的疾病之一，引起人類瘧疾的所有瘧原蟲都是按蚊屬昆蟲傳播的，，非洲岡比亞按蚊高度嗜吸人血，是惡性瘧原蟲極其高效的傳播媒介，所以危害尤為嚴(yán)重，昆蟲都是依靠嗅覺發(fā)現(xiàn)寄主的，嗅覺在按蚊的搜索寄主行為中起關(guān)鍵作用，昆蟲能夠感受空氣中的揮發(fā)性氣味分子，并依此作為尋偶、覓食和尋找產(chǎn)卵場(chǎng)所的信息。而昆蟲觸角感受器中的氣味結(jié)合蛋白被認(rèn)為是運(yùn)輸氣味分子的受體，是昆蟲專一性識(shí)別外界氣味物質(zhì)的第一步生化反應(yīng)，對(duì)于昆蟲與外界進(jìn)行信息交流具有重要意義，本論文開展了對(duì)岡比亞按蚊的氣味結(jié)合蛋白AgOBP48基因的研究。 利用英國(guó)洛桑研究所惠贈(zèng)的含有AgOBP48基因的PET-20b(+)，用NdeⅠ／EcoRⅠ雙酶切取得目的基因片段，將目的片斷和原核表達(dá)載體pET-28a(+)經(jīng)NdeⅠ／EcoRⅠ雙酶酶切重組到表達(dá)型質(zhì)粒中并轉(zhuǎn)化入原核細(xì)胞中。表達(dá)序列測(cè)定結(jié)果表明：岡比亞按蚊氣味結(jié)基因AgOBP48的成熟蛋白閱讀框全長(zhǎng)536bp，編碼172個(gè)氨基酸殘基，預(yù)測(cè)分子量和等電點(diǎn)分別為19kDa和5.00。 本研究將岡比亞按蚊AgOBP48基因整合入原核表達(dá)載體中表達(dá)，得到的重組蛋白在其N-端融合有His·Tag純化標(biāo)記，為下一步蛋白的純化，功能研究奠定基礎(chǔ)。
[Abstract]:Malaria is one of the diseases that seriously endanger human health, and all malaria parasites causing human malaria are transmitted by Anopheles mosquitoes, and the African Anopheles gambiae are highly blood-sucking and extremely efficient vectors of transmission of Plasmodium falciparum, So the damage is especially serious. Insects rely on smell to find their hosts. Smell plays a key role in the search for the host of Anopheles. Insects can sense volatile odour molecules in the air and use it as a match. Information about foraging and finding spawning places. The odor binding protein in the antennae is considered to be the receptor for the transport of odour molecules and the first step biochemical reaction for insect to specifically recognize the external odour substances. It is of great significance for the communication of information between insects and the outside world. In this paper, we studied the AgOBP48 gene of odour binding protein in Anopheles gambiae (Anopheles gambiae). The target gene fragment was obtained by using Nde 鈪

本文編號(hào)：2296193