Study On Blood Transcripts Of Wolves And Dogs Based On High - Throughput Sequencing

Both of wolf(Canis lupus) and dog(Canis lupus familiaris) are classified under order Carnivora, family Canidae, genus Canis. Because of their similar appearance, the study of their evolutionary relationships has always been a hot area. But so far, about their research was mainly concentrated in the mitochondria and genome. The dog’s genome has been assembly and add to public database for researchers to use free of charge by the national human genome research institute(NHGRI) in 2004. We analysis blood transcriptome of wolves and dogs based on the dog’s genome. The following results were obtained:1. Based on RNA- Seq technology, through the two dogs and three Wolf blood transcriptome sequencing, after filtering a total of 5.33 G, 6.24 G, 6.4 G, 6.28 G and 6 G of high-quality reads was obtain from 30.87 G row reads, about 97% of the total. With reference to dog genome, more than 85% of each reads map to reference genome. Of the total reads, more than 60% reads was uniquely mapped reads, while less than 30% reads was multiple mapped reads. After the assemble, we obtained 26312 genes, including 1989 novel genes and identified a total of 33229 transcripts, including 1993 novel transcripts.2. Based on the results of the Cufflinks, we screening of differentially expressed genes by DESeq. After setting threshold p-value < 0.05, a total of 524 differentially expressed genes was identified by comparison and manual annotation with database. Of these genes, 180 up-regulated genes including 12 novel genes and 344 down-regulated genes including 23 novel genes. In these differentially expressed genes, There are three genes are expressed only in the Wolf, seven gene is expressed only in the dog in these differentially expressed genes.3. We had the GO enrichment analysis to find the biological processes of these differentially expressed genes. According to the results, a total of 272 genes were enriched to 703 GO terms, 522 in molecular function, 100 in biological process, 81 in cellular component. After setting threshold p-value < 0.05, all of the differentially expressed genes have been significantly enriched in 382 GO terms, accounted for 54% of all the terms. Then in order to understand the wolves and dogs differentially expressed genes involved in biological processes more clearly, we had GO enrichment analysis of their high expression differentially genes respective. It was found that the Wolf has six genes was enriched on the wolf’s innate immune response.4. KEGG pathway analysis results show that the enrichment of a total of 218 genes on the 12 KEGG pathways, including cell cycle, Parkinson’s disease, oxidative phosphorylation, protein process in endoplasmic reticulum, and p53 signaling pathway and so on.This study had transcriptome sequencing and analysis of the wolf and dog’s blood for the first time. To further explore the evolutionary relationships of wolves and dogs, and provide blood transcription data to reveal the reason for the difference between wolf and dog in blood immune.