中华按蚊八个羧肽酶基因的鉴定及其与吸血餐相关的表达分析（英文）

发布时间：2018-03-27 18:19

  本文选题：中华按蚊　切入点：羧肽酶　出处：《昆虫学报》2017年06期

【摘要】：【目的】本研究旨在鉴定中华按蚊Anopheles sinensis 8个消化型羧肽酶基因,并分析这些羧肽酶基因在血餐前后的表达模式。【方法】通过生物信息学方法分析中华按蚊8个羧肽酶基因的特征,采用半定量PCR或定量PCR(qRT-PCR)方法分别分析这些基因在中华按蚊不同发育时期不同组织和在取食血液前后中华按蚊雌成虫中肠中的表达情况。【结果】通过生物信息学分析发现,8个羧肽酶基因中,6个编码羧肽酶A(AsCPA-I~AsCPA-VI),2个编码羧肽酶B(AsCPB-I和AsCPB-II)。表达分析发现,只有AsCPA-V在中华按蚊4龄幼虫中肠和中肠外组织(仅去掉中肠的虫体)中表达,可能为幼虫特异性表达的基因,其余7个羧肽酶基因既在幼虫期表达又在成虫期表达。取食血液后,AsCPA-I,AsCPA-II,AsCPA-III,AsCPA-IV,AsCPA-VI,AsCPB-I和AsCPB-II在中华按蚊雌成虫中肠中的表达明显受到血液的影响,但表达模式不一样,说明血液蛋白的消化是由多个基因协同作用的复杂生理过程。其中,AsCPA-I,AsCPA-III,AsCPA-IV,AsCPA-VI和AsCPB-II在取食血液后表达上调,且都在吸血后的24 h达到表达高峰。尤其是AsCPA-VI在吸血后24 h表达上调了约418倍,AsCPB-II上调了40倍,可能与血液蛋白的消化有关。而AsCPA-II和AsCPB-I基因的表达水平在中华按蚊取食血液后显著下调。【结论】本研究对中华按蚊8个消化型羧肽酶基因进行了鉴定和表达分析。AsCPA-I,AsCPA-III,AsCPA-IV,AsCPA-VI和AsCPB-II可能参与血液蛋白的消化,尤其是AsCPA-VI和AsCPB-II可能起着更为重要的作用。研究结果为深入剖析中华按蚊血液蛋白消化的分子机理奠定了基础。
[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.
【作者单位】： 重庆师范大学昆虫与分子生物学研究所重庆市媒介昆虫重点实验室;
【基金】：国家自然科学基金项目(31572332,31672363) 重庆市教委科学技术研究项目(KJ1400507) 重庆市优秀人才支持计划 “两江学者”计划专项经费资助项目
【分类号】：Q966

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