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通过RNA测序进行比较转录组分析,以识别成年伯克夏猪和济州本地猪肝脏和肌肉组织中基因的差异表达。

Comparative transcriptomic analysis by RNA-seq to discern differential expression of genes in liver and muscle tissues of adult Berkshire and Jeju Native Pig.

作者信息

Sodhi Simrinder Singh, Song Ki-Duk, Ghosh Mrinmoy, Sharma Neelesh, Lee Sung Jin, Kim Jeong Hyun, Kim Nameun, Mongre Raj Kumar, Adhikari Pradeep, Kim Jin Young, Hong Sang Pyo, Oh Sung Jong, Jeong Dong Kee

机构信息

Department of Animal Biotechnology, Faculty of Biotechnology, Jeju National University, Jeju 690-756, South Korea.

The Animal Genomics and Breeding Center, Hankyong National University, Anseong-si, Gyeonggi-do 456-749, South Korea.

出版信息

Gene. 2014 Aug 10;546(2):233-42. doi: 10.1016/j.gene.2014.06.005. Epub 2014 Jun 6.

Abstract

RNA-seq is being rapidly adopted for the profiling of the transcriptomes in different areas of biology, especially in the studies related to gene regulation. The discovery of differentially expressed genes (DEGs) between adult animals of Jeju Native Pig (JNP) and Berkshire breeds of Sus scrofa, is of particular interest for the current study. For the better understanding of the gene expression profiles of the liver and longissimus dorsi muscle, DEGs were identified via RNA-seq. Sequence reads were obtained from Illumina HiSeq2000 and mapped to the pig reference genome (Sscrofa10.2) using Tophat2. We identified 169 and 39 DEGs in the liver and muscle of JNP respectively, by comparison with Berkshire breed. Out of all identified genes, 41 genes in the liver and 9 genes in the muscle have given significant expression. Gene ontology (GO) terms of developmental process and KEGG pathway analysis showed that metabolic, immune response and protein binding were commonly enriched pathways in the two tissues. Further the heat map analysis by ArrayStar has shown the different levels of expression in JNP with respect to the Berkshire breed. The validation through real time PCR and western blotting also confirmed the differential expression of genes in both breeds. Genes pertaining to metabolic process and inflammatory and immune system are more enriched in Berkshire breed. This comparative transcriptome analysis of two tissues suggests a subset of novel marker genes which expressed differently between the JNP and Berkshire.

摘要

RNA测序正迅速应用于生物学不同领域的转录组分析,尤其是在与基因调控相关的研究中。济州本地猪(JNP)成年动物与家猪伯克夏品种之间差异表达基因(DEG)的发现,是本研究特别感兴趣的内容。为了更好地了解肝脏和背最长肌的基因表达谱,通过RNA测序鉴定了差异表达基因。序列读数从Illumina HiSeq2000获得,并使用Tophat2映射到家猪参考基因组(Sscrofa10.2)。通过与伯克夏品种比较,我们在JNP的肝脏和肌肉中分别鉴定出169个和39个差异表达基因。在所有鉴定出的基因中,肝脏中的41个基因和肌肉中的9个基因有显著表达。基因本体(GO)术语的发育过程和KEGG通路分析表明,代谢、免疫反应和蛋白质结合是这两个组织中常见的富集通路。进一步通过ArrayStar进行的热图分析显示了JNP相对于伯克夏品种的不同表达水平。通过实时PCR和蛋白质印迹法进行的验证也证实了两个品种中基因的差异表达。与代谢过程以及炎症和免疫系统相关的基因在伯克夏品种中更为富集。这两个组织的比较转录组分析表明了一组新的标记基因,它们在JNP和伯克夏之间表达不同。

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