基于GO分析的全基因组芯片筛选小鼠牙囊发育早期差异表达基因的研究
Screen of the differentially expressed genes in early developing murine dental follicle tissue based on GO analysis with whole genome microarray
张春光 1聂敏媛 2孟庆岩 2王洪云 3安钰 4董蕊 5党平2
作者信息
- 1. 解放军第323医院,陕西 西安 710054
- 2. 解放军第306医院,北京 100101
- 3. 天津海滨人民医院口腔科,天津 300280
- 4. 北大国际医院,北京 102206
- 5. 首都医科大学口腔医学院,北京 100050
- 折叠
摘要
目的:利用基于GO 分析的全基因组芯片技术筛选小鼠牙囊组织在个体发育不同时期的差异表达基因.方法:以激光显微切割技术获取胚胎17 d(E17)和出生后2 d(PN2)两个时期小鼠下颌第一磨牙的牙囊组织后,以改良Trizol法提取总RNA并制备荧光标记的cRNA;然后再以Aglient 小鼠全基因组表达芯片对比分析上述两时期的基因表达差异;最后以GeneSpringGX 7.3软件对所得结果进行归一化及倍数筛选,并分别以BioCarta、GenMAPP软件进行基因分类和GO分析.结果:①得到两个发育时期牙囊组织相对于牙乳头组织表达倍数都在10倍以上的一组基因;②得到细胞外基质等10余项常见生物功能相关的基因列表.结论:将激光显微切割以及基因表达芯片两种技术联合可用于小鼠牙周组织发育过程中差异表达基因.
Abstract
AIM:To screen the differentially expressed genes in early developing murine dental follicle tissue by whole genome microarray based on GO analysis.METHODS:The murine dental follicle at E17 and PN2 stages were dissected by LMD6000 system (Laser Micro Dissected system,Leica).To ensure the purity and the integrity of RNA samples,quality check was adopted by Bioanalyzer 2100 System,Agilent.For control and test RNAs,the synthesis of target cRNA probes and hybridization were performed using Agilent's Low RNA Input Linear Amplification kit.The fragmented cRNA was hybridized with assembled Agilent's Mouse Oligo Microarray.All microarray data normalization and selection of fold-changed genes were performed using GeneSpringGX 7.3.Functional annotation of the genes and GO analysis werer performed according to Gene OntologyTM Consortium by GeneSpringGX 7.3.Gene classification was based on searches by BioCarta,GenMAPP.RESULTS:A group of genes which were highly expressed in dental follicle compared to dental papilla tissue at both stages were obtained.More than 10 genes related to common biological functions such as "extracellular matrix" were obtained.CONCLUSION:The combination of laser microdissection and Microarray / bioinformatics analysis is a valuable means to obtain a global view of gene expression during the murine root / periodontal tissue development process.
关键词
激光显微切割技术/牙胚发育/基因芯片/差异表达基因Key words
laser microdissection/tooth germ development/microarray/differential expression of genes引用本文复制引用
基金项目
首都卫生发展科研专项(首发2014-4-5051)
解放军第306医院院级科题(13QN04)
出版年
2017
NETL
NSTL
维普
万方数据