淡色库蚊氨肽酶N基因在不同龄期幼虫的表达谱分析

doi:10.11853/j.issn.1003.8280.2016.05.007

中国媒介生物学及控制杂志 ›› 2016, Vol. 27 ›› Issue (5): 450-453.DOI: 10.11853/j.issn.1003.8280.2016.05.007

淡色库蚊氨肽酶N基因在不同龄期幼虫的表达谱分析

郭秀霞, 杨培培, 程鹏, 王海防, 刘丽娟, 张崇星, 王怀位, 公茂庆

山东省医学科学院, 山东省寄生虫病防治研究所, 山东省医药卫生媒介生物学重点实验室, 山东济宁 272033

收稿日期:2016-06-16 出版日期:2016-10-20 发布日期:2016-10-20
通讯作者: 公茂庆,Email:maoqingg@yahoo.com
作者简介:郭秀霞,女,硕士,助理研究员,从事病原生物学研究,Email:guoguoxiua@163.com
基金资助:
国家自然科学基金（81271877）

Expression profile of aminopeptidase N in different instars of Culex pipiens pallens larvae

GUO Xiu-xia, YANG Pei-pei, CHENG Peng, WANG Hai-fang, LIU Li-juan, ZHANG Chong-xing, WANG Huai-wei, GONG Mao-qing

Shandong Institute of Parasitic Diseases, Shandong Academy of Medical Sciences, Key Laboratory of Medicine and Health in Shandong Province, Jining 272033, Shandong Province, China

Received:2016-06-16 Online:2016-10-20 Published:2016-10-20
Supported by:
Supported by the National Natural Science Foundation of China (No. 81271877)

摘要/Abstract

摘要：

目的对淡色库蚊不同龄期幼虫中肠氨肽酶N（Aminopeptidase N，APN）的表达水平进行分析。方法应用反转录-聚合酶链反应（RT-PCR）克隆淡色库蚊APN基因的全长cDNA序列，利用实时荧光定量PCR（qRT-PCR）分析该基因在不同龄期幼虫中肠的表达情况。结果克隆获得淡色库蚊APN基因，该基因全长1 383 bp，编码460个氨基酸，与GenBank收录的致倦库蚊（XM001862270.1）序列的同源性为100%。qRT-PCR表明CpAPN基因在各个龄期均有表达，其中在4龄期的表达量最高，3龄期最低，1、2和4龄期的表达量差异无统计学意义（F=2.465，P=0.250），与3龄期幼虫的差异有统计学意义（F=49.191，P=0.000），分别为其表达量的3.5、3.7和4.0倍。结论 CpAPN基因在淡色库蚊不同龄期幼虫中的表达水平存在差异，为进一步探讨淡色库蚊APN的功能奠定了基础。

关键词: 淡色库蚊, 氨肽酶N, 龄期, 幼虫, 表达谱

Abstract:

Objective To analyze the expression levels of CpAPN in different instars of Culex pipiens pallens. Methods The APN cDNA sequence was identified and sequenced by quantitative Real-time PCR (qRT-PCR), and the relative expression levels of CpAPN were assessed in different instars of larvae using qRT-PCR assays. Results The full-length of CpAPN was 1 383 bp that encoded a protein of 460 amino acid residues. The homology with Cx. pipiens quinquefasciatus (XM001862270.1) was 100%. The qRT-PCR showed that APN was expressed at all stages of Cx. pipiens pallens, with the highest level in the 4^th instar and the lowest level in the 3^rd instar. There was no significant difference among the 1^st, 2^nd and 4^th instars (F=2.465,P=0.250), but they were all higher than the 3^rd instar significantly (F=49.191, P=0.000), which were 3.5 times, 3.7 times, and 4.0 times of the expression levels to the 3^rd instar, respectively. Conclusion CpAPN showed significantly different expression levels in different instars of Cx. pipiens pallens. The results provided experimental evidences for revealing the function of CpAPN of the mosquitoes.

Key words: Culex pipiens pallens, Aminopeptidase N, Instar, Larvae, Expression profiling

中图分类号:

R384.1

郭秀霞, 杨培培, 程鹏, 王海防, 刘丽娟, 张崇星, 王怀位, 公茂庆. 淡色库蚊氨肽酶N基因在不同龄期幼虫的表达谱分析[J]. 中国媒介生物学及控制杂志, 2016, 27(5): 450-453.

GUO Xiu-xia, YANG Pei-pei, CHENG Peng, WANG Hai-fang, LIU Li-juan, ZHANG Chong-xing, WANG Huai-wei, GONG Mao-qing. Expression profile of aminopeptidase N in different instars of Culex pipiens pallens larvae[J]. Chines Journal of Vector Biology and Control, 2016, 27(5): 450-453.

参考文献

[1] Gemperli A, Sogoba N, Fondjo E, et al. Mapping malaria transmission in West and Central Africa[J]. Trop Med Int Health,2006,1(7):1032-1046.
[2] Onyango SA, Kitron U, Mungai P, et al. Monitoring malaria vector control interventions: effectiveness of five different adult mosquito sampling methods[J]. J Med Entomol,2013,50(5): 1140-1151.
[3] Bravo A, Likitvivatanavong S, Gill SS, et al. Bacillus thuringiensis: a story of a successful bioinsecticide[J]. Insect Biochem Mol Biol,2011,41(7):423-431.
[4] Vachon V, Laprade R, Schwartz JL. Current models of the mode of action of Bacillus thuringiensis insecticidal crystal proteins: a critical review[J]. J Invertebr Pathol,2012,111(1):1-12.
[5] Ren XL, Ma Y, Cui JJ, et al. RNA interference-mediated knockdown of three putative aminopeptidases N affects susceptibility of Spodoptera exigua larvae to Bacillus thuringiensis Cry1Ca[J]. J Insect Physiol,2014,67:28-36.
[6] Pigott CR, Ellar DJ. Role of receptors in Bacillus thuringiensis Crystal toxin activity[J]. Microbiol Mol Biol Rev,2007,71(2): 255-281.
[7] Crava CM, Bel Y, Lee SF, et al. Study of the aminopeptidase N gene family in the lepidopterans Ostrinia nubilalis (Hübner) and Bombyx mori (L.): sequences, mapping and expression[J]. Insect Biochem Mol Biol,2010,40(7):506-515.
[8] Denolf P, Hendrickx K, Van Damme J, et al. Cloning and characterization of Manduca sexta and Plutella xylostella midgut aminopeptidase N enzymes related to Bacillus thuringiensis toxin-binding proteins[J]. Eur J Biochem,1997,248(3):748-761.
[9] Chen JW, Aimanova KG, Pan SQ, et al. Identification and characterization of Aedes aegypti aminopeptidase N as a putative receptor of Bacillus thuringiensis Cry11A toxin[J]. Insect Biochem Mol Biol,2009,39(10):688-696.
[10] Rajagopal R,Agrawal N,Selvapandiyan A, et al. Recombinantly expressed isoenzymic aminopeptidases from Helicoverpa armigera (American cotton bollworm) midgut display differential interaction with closely related Bacillus thuringiensis insecticidal proteins[J]. Biochem J,2003,370(3):971-978.
[11] Aroonkesorn A, Pootanakit K, Katzenmeier G, et al. Two specific membrane-bound aminopeptidase N isoforms from Aedes aegypti larvae serve as functional receptors for the Bacillus thuringiensis Cry4Ba toxin implicating counterpart specificity[J]. Biochem Biophys Res Commun,2015,461(2):300-306.
[12] Angelucci C, Barrett-Wilt GA, Hunt DF, et al. Diversity of aminopeptidases, derived from four lepidopteran gene duplications, and polycalins expressed in the midgut of Helicoverpa armigera: identification of proteins binding the δ-endotoxin, Cry1Ac of Bacillus thuringiensis[J]. Insect Biochem Mol Biol,2008,38(7):685-696.

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淡色库蚊氨肽酶N基因在不同龄期幼虫的表达谱分析

Expression profile of aminopeptidase N in different instars of Culex pipiens pallens larvae

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