本文選題：中華按蚊　切入點(diǎn)：羧肽酶　出處：《昆蟲學(xué)報(bào)》2017年06期

【摘要】：【目的】本研究旨在鑒定中華按蚊Anopheles sinensis 8個(gè)消化型羧肽酶基因,并分析這些羧肽酶基因在血餐前后的表達(dá)模式�！痉椒ā客ㄟ^生物信息學(xué)方法分析中華按蚊8個(gè)羧肽酶基因的特征,采用半定量PCR或定量PCR(qRT-PCR)方法分別分析這些基因在中華按蚊不同發(fā)育時(shí)期不同組織和在取食血液前后中華按蚊雌成蟲中腸中的表達(dá)情況�！窘Y(jié)果】通過生物信息學(xué)分析發(fā)現(xiàn),8個(gè)羧肽酶基因中,6個(gè)編碼羧肽酶A(AsCPA-I~AsCPA-VI),2個(gè)編碼羧肽酶B(AsCPB-I和AsCPB-II)。表達(dá)分析發(fā)現(xiàn),只有AsCPA-V在中華按蚊4齡幼蟲中腸和中腸外組織(僅去掉中腸的蟲體)中表達(dá),可能為幼蟲特異性表達(dá)的基因,其余7個(gè)羧肽酶基因既在幼蟲期表達(dá)又在成蟲期表達(dá)。取食血液后,AsCPA-I,AsCPA-II,AsCPA-III,AsCPA-IV,AsCPA-VI,AsCPB-I和AsCPB-II在中華按蚊雌成蟲中腸中的表達(dá)明顯受到血液的影響,但表達(dá)模式不一樣,說明血液蛋白的消化是由多個(gè)基因協(xié)同作用的復(fù)雜生理過程。其中,AsCPA-I,AsCPA-III,AsCPA-IV,AsCPA-VI和AsCPB-II在取食血液后表達(dá)上調(diào),且都在吸血后的24 h達(dá)到表達(dá)高峰。尤其是AsCPA-VI在吸血后24 h表達(dá)上調(diào)了約418倍,AsCPB-II上調(diào)了40倍,可能與血液蛋白的消化有關(guān)。而AsCPA-II和AsCPB-I基因的表達(dá)水平在中華按蚊取食血液后顯著下調(diào)。【結(jié)論】本研究對(duì)中華按蚊8個(gè)消化型羧肽酶基因進(jìn)行了鑒定和表達(dá)分析。AsCPA-I,AsCPA-III,AsCPA-IV,AsCPA-VI和AsCPB-II可能參與血液蛋白的消化,尤其是AsCPA-VI和AsCPB-II可能起著更為重要的作用。研究結(jié)果為深入剖析中華按蚊血液蛋白消化的分子機(jī)理奠定了基礎(chǔ)。
[Abstract]:[objective] to identify eight digestive carboxypeptidase genes of Anopheles sinensis Anopheles sinensis. The expression patterns of these carboxypeptidase genes before and after blood meal were analyzed. [methods] the characteristics of 8 carboxypeptidase genes in Anopheles sinensis were analyzed by bioinformatics. The expression of these genes in the midgut of Anopheles sinensis at different developmental stages and before and after feeding blood was analyzed by semi-quantitative PCR or quantitative PCRQRT-PCR. [results] Bioinformatics was used to analyze the expression of these genes in the midgut of Anopheles sinensis female adults. Six of the 8 carboxypeptidase genes were identified as ASCPA-VIN, 2 encoding carboxypeptidase B(AsCPB-I and 2 ASCPB-III. Only AsCPA-V was expressed in the midgut and parenteral tissue of Anopheles sinensis 4th instar larvae (only the midgut body was removed), which may be a specific larval expression gene. The other 7 carboxypeptidase genes were expressed in both larval and adult stages. The expression of AsCPA-Ittrium AsCPA-IIIA- AsCPA-IVA- AsCPA-VIPB-I and AsCPB-II in the midgut of female adult Anopheles sinensis was significantly affected by the blood, but the expression pattern was not the same, but the expression pattern was different from that of Anopheles sinensis (Anopheles sinensis). These results indicate that the digestion of blood protein is a complex physiological process by the synergistic action of multiple genes, and the expression of AsCPA-IIIA- AsCPA-IVP AsCPA-VI and AsCPB-II is up-regulated after feeding blood. The expression of AsCPA-VI was increased about 418 times and 40 times of AsCPB-II at 24 h after blood sucking, and the expression of ASCPB-II was up to 40 times higher than that of ASCPB-II. The expression levels of AsCPA-II and AsCPB-I genes were significantly down-regulated by Anopheles sinensis after feeding on blood. [conclusion] eight digestive carboxypeptidase genes of Anopheles sinensis were identified and expressed in this study. AsCPA-Ittrium AsCPA-IIIN AsCPA-IVAVI and AsCPB-II may be involved in the digestion of blood proteins, In particular, AsCPA-VI and AsCPB-II may play a more important role. The results laid a foundation for further analysis of the molecular mechanism of Anopheles sinensis blood protein digestion.
【作者單位】： 重慶師范大學(xué)昆蟲與分子生物學(xué)研究所重慶市媒介昆蟲重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金項(xiàng)目(31572332,31672363) 重慶市教委科學(xué)技術(shù)研究項(xiàng)目(KJ1400507) 重慶市優(yōu)秀人才支持計(jì)劃 “兩江學(xué)者”計(jì)劃專項(xiàng)經(jīng)費(fèi)資助項(xiàng)目
【分類號(hào)】：Q966

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本文編號(hào)：1672683