棕尾别麻蝇蛹3个发育时期的转录组差异分析

中国寄生虫学与寄生虫病杂志 ›› 2017, Vol. 35 ›› Issue (3): 274-279.

棕尾别麻蝇蛹3个发育时期的转录组差异分析

周昊, 王启燕, 张红玲, 任峥, 戴佳琳, 刘玉波, 王杰, 黄江^*()

贵州医科大学法医学院,法医司法鉴定中心,贵阳 550004

收稿日期:2016-11-30 出版日期:2017-03-30 发布日期:2017-09-07
通讯作者: 黄江
基金资助:
国家自然科学基金（No. 81641082）;贵州省高校工程技术研究中心项目（黔科合KY字［2013］114号）;贵州省现代法医司法鉴定工程研究中心（黔发改高技［2016］1345）;贵州省科技厅计划基金（黔科合LH字［2016］7360）

Differential analysis of transcriptomes in Boettcherisca peregrine pupae at three developmental stages

Hao ZHOU, Qi-yan WANG, Hong-ling ZHANG, Zheng REN, Jia-lin DAI, Yu-bo LIU, Jie WANG, Jiang HUANG^*()

School of Forensic Medicine, Guizhou Medical University, Guiyang 550004, China

Received:2016-11-30 Online:2017-03-30 Published:2017-09-07
Contact: Jiang HUANG
Supported by:
Supported by the National Natural Science Foundation of China(No. 81641082);the Guizhou University Engineering Technology Research Center Project(Qian Science KY No.［2013］114);the Guizhou Modern Forensic Judicial Authentication Engineering Research Center(Qian Science No.［2016］1345);and Project Research Foundation of the Science and Technology Department of Guizhou province(Qian Science LH No.［2016］7360)

摘要/Abstract

摘要： 目的探讨棕尾别麻蝇（Boettcherisca peregrine）蛹3个时期转录组的差异表达,为进一步探究棕尾别麻蝇蛹各个时期转录组变化提供实验数据。方法提取棕尾别麻蝇蛹3个时期第1（MY1）、5（MY2）和10天（MY3）的蝇蛹总RNA,反转录成cDNA,并进行测序,使用软件edgeR对基因差异表达进行分析,对差异基因进行GO分类统计和富集分析;对已被注释的基因单基因簇（Unigene）进行KEGG代谢通路分析;使用MISA对Unigene进行简单重复序列（SSR）检测。结果蝇蛹3个时期cDNA经基因差异表达分析,得到各时期样品基因组表达水平,共捕获38 727条Unigenes,将不同时期的蝇蛹转录组进行两两比较,其差异表达基因数介于1 263~2 430,差异基因总数为5 283。聚类图型得到MY1、MY2和MY3组的两个样品表达模式相似,聚为一支。棕尾别麻蝇蛹转录组共有8 856个Unigenes被注释,被注释到的代谢通路有336个。SSR查找发现,从8 645个Unigenes中共查找到12 146个SSR位点,占Unigene总数的22.3%。其中,单核苷酸重复所占比例最高,达到69.2%,其次是三核苷酸重复（21.0%）。SSR不同重复基元类型中,出现频率最高的为A/T,其次是AAC/GTT、AG/GT。结论棕尾别麻蝇蛹在不同时期基因表达有显著差异.

关键词: 棕尾别麻蝇, 蛹, 转录组, 基因差异表达

Abstract: Objective To investigate the differential expression of transcriptomes in Boettcherisca peregrine pupae at three developmental stages. Methods Total RNA was extracted from B. peregrine pupae at stages of day 1(MY1), day 5(MY2) and day 10(MY3), reverse-transcribed into cDNA, and sequenced. Gene differential expression was analyzed with edgeR software. GO enrichment analysis was made for genes with differential expression. Analysis of KEGG(Kyoto Encyclopedia of Genes and Genomes) metabolic pathway was made for Universal Gene (Unigene). The simple sequence repeat(SSR) of Unigene was detected with MISA. Results A total of 38 727 Unigenes were obtained after differential expression analysis on cDNA at the three stages. The numbers of genes with differential expression between each two stages were 1 263-2 430, with an overall number of 5 283. Pattern clustering analysis revealed similar expression patterns of those genes at the three stages. A total of 8 856 Unigenes and 336 pathways were annotated for B. peregrine pupae. A total of 12 146 SSR were detected from 8 645 Unigenes(22.3%), comprising predominantly of single-nucleotide repeats(69.23%), then the three-nucleotide repeat(21.0%). Of the SSR, the A/T occurred most frequently, followed by AAC/GTT and AG/GT. Conclusions There are significant differences in gene expression in B. peregrine pupae at different developmental stages.

Key words: Boettcherisca peregrine, Pupa, Transcriptome, Differential expression of genes

中图分类号:

R384.21

周昊, 王启燕, 张红玲, 任峥, 戴佳琳, 刘玉波, 王杰, 黄江. 棕尾别麻蝇蛹3个发育时期的转录组差异分析[J]. 中国寄生虫学与寄生虫病杂志, 2017, 35(3): 274-279.

Hao ZHOU, Qi-yan WANG, Hong-ling ZHANG, Zheng REN, Jia-lin DAI, Yu-bo LIU, Jie WANG, Jiang HUANG. Differential analysis of transcriptomes in Boettcherisca peregrine pupae at three developmental stages[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2017, 35(3): 274-279.

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基因名称 Gene name	引物名 Primer name	引物序列（5′→ 3′） Primer sequences（5′→ 3′）	长度/bp Length/bp	退火温度/℃ Tm/℃
TH	TH-F： TH-R：	ATGGTGCTGGTCTCTTGTCC ATACCCAACGACGGAACTTG	3 625	60
CBM-14	CBM-14-F： CBM-14-R：	AAACACCACGGAAATCATCC CCAGAAGCAATCATCCCAAT	663	60
Tubulin	Tubulin-F： Tubulin-R：	GCTTCCTCATCTTCCACTCGTTC GATGTCGTAGATAGCCTCGTTGTC	1 353	60

代谢通路ID Metabolic pathway ID	代谢通路 Metabolic pathway	被注释的数量 Annotated number	所占比例/% Percentage/%
ko01100	生化代谢Metabolic pathways	1 353	15.28
ko01110	次生代谢产物的生物合成Biosynthesis of secondary metabolites	438	4.95
ko05016	亨廷顿氏舞蹈症Huntington's disease	405	4.57
ko01120	微生物的代谢Microbial metabolism in	273	3.08
ko05010	阿尔茨海默病Alzheimer's disease	254	2.87
ko05200	恶性肿瘤通路Alzheimer's disease	229	2.59
ko00230	嘌呤代谢Purine metabolism	214	2.42
ko00190	氧化磷酸化Oxidative phosphorylation	212	2.39
ko05012	帕金森病Parkinson's disease	199	2.25
ko05166	HTLV-I感染HTLV-I infection	198	2.24

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