猪卵母细胞GV/MⅡ期mRNA/lncRNA差异表达的研究Differential Expression of GV/MⅡ Phase mRNA/lncRNA in Porcine Oocytes
焦亚飞;刘树林;叶升;高邦君;石德顺;黄奔;
JIAO Yafei;LIU Shulin;YE Sheng;Gao Bangjun;SHI Deshun;HUANG Ben;State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, College of Animal Science and Technology, Guangxi University;
摘要(Abstract):
为解决体外成熟的猪卵母细胞受精后形成原核率低、多精入卵率高及体外成熟的卵母细胞囊胚率低等问题,本实验以猪卵母细胞为材料,采用转录组学的方法探讨卵母细胞成熟过程中mRNA/lncRNA的表达差异,筛选出影响猪卵母细胞成熟的关键mRNA/lncRNA。结果表明:本研究构建了2个时期的RNA文库,2组文库一共检测到已知的mRNA 1 753 030个,其中共表达mRNA16 469个,2 486个mRNA差异显著,其中上调基因752个,下调基因1 734个;此次转录组测序共鉴定出lncRNA 22 811个,2个时期差异表达已知lncRNA 15个,通过随机挑选10个测序所得到的mRNA转录本和12个lncRNA转录本进行QRT-PCR验证,基本与高通量测序保持一致,说明此次转录组测序结果符合要求,数据准确可靠,可以用于后续分析研究。
In order to solve the problems of low prokaryotic formation rate, high yield of polyspermy and low blastulation rate after fertilization of porcine oocytes in vitro, we need to understand the related mechanism of oocyte maturation in vitro. The transcriptomics method was used to investigate the differential expression of mRNA/lncRNA during oocyte maturation, and the key mRNA/lncRNA affecting the maturation of porcine oocytes was screened to lay a foundation for the study of oocyte maturation mechanism. The results showed that two RNA libraries were constructed in this study, a total of 1 753 030 known mRNAs were detected in the two groups, a total of 16,469 mRNAs were co-expressed, and 2,486 mRNAs were significantly different, of which 752 were up-regulated and 1,734 were down-regulated. In addition, a total of 22,811 lncRNAs were identified by transcriptome sequencing, two periods of differential expression of known 15 lncRNAs. The 10 mRNA transcripts and 12 lncRNA transcripts obtained through random selection were verified by QRT-PCR, the results showed that they were basically consistent with high-throughput sequencing, indicating that the transcriptome sequencing results met the requirements, and the data is accurate and reliable, and could be used for subsequent analysis and research.
关键词(KeyWords):
猪;卵母细胞;mRNA;lncRNA;转录组测序
Porcine;Oocyte;mRNA;LncRNA;Transcriptome sequencing
基金项目(Foundation): 国家自然科学基金资助项目(31401267)
作者(Authors):
焦亚飞;刘树林;叶升;高邦君;石德顺;黄奔;
JIAO Yafei;LIU Shulin;YE Sheng;Gao Bangjun;SHI Deshun;HUANG Ben;State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, College of Animal Science and Technology, Guangxi University;
DOI: 10.19556/j.0258-7033.20190326-14
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- 焦亚飞
- 刘树林
- 叶升
- 高邦君
- 石德顺
- 黄奔
JIAO Yafei- LIU Shulin
- YE Sheng
- Gao Bangjun
- SHI Deshun
- HUANG Ben
- State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources
- College of Animal Science and Technology
- Guangxi University
- 焦亚飞
- 刘树林
- 叶升
- 高邦君
- 石德顺
- 黄奔
JIAO Yafei- LIU Shulin
- YE Sheng
- Gao Bangjun
- SHI Deshun
- HUANG Ben
- State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources
- College of Animal Science and Technology
- Guangxi University