ISSN 1003-8280 CN 10-1522/R 中国疾病预防控制中心 主办
收稿日期: 2016-06-16
网络出版日期: 2016-10-20
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
Received date: 2016-06-16
Online published: 2016-10-20
Supported by
Supported by the National Natural Science Foundation of China (No. 81271877)
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。
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 . DOI: 10.11853/j.issn.1003.8280.2016.05.007
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 4th instar and the lowest level in the 3rd instar. There was no significant difference among the 1st, 2nd and 4th instars (F=2.465,P=0.250), but they were all higher than the 3rd 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 3rd 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
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