A Preliminary Study On Immunobiology Of Porcine DNMT1 Gene

Epigenetics, with different Chinese translated names such as “拟遗传学”, “表遗传学”, “外遗传学”, and “后遗传学”, is an important part of the soft inheritance. Although epigenetics involves multiple aspects of inheritance, DNA methylation is one of the most core and important part of the epigenetic studies. DNMT1 is a kind of methylation transferase, mainly performing the biological function of DNA methylation maintenance in the cell. At present, a lot of researches on DNMT1 gene were carried out in plants and animals such as human, mice, and arabidopsis thaliana, and a series of fruitful results have been obtained. However, these researches mainly concentrated on model animals and plants, while there were not enough in-depth investigations in livestock, and systematical exploration of DNMT1 gene function from the immune view is more insufficient.DNMT1 gene is involved in the regulation of the most biological processes within the body, and it also has close relation with immune regulation. In view of this, this study uses pig as the research object, and mainly focuses on immunity, intending to do preliminary research on the pig DNMT1 gene, respectively from the aspects of the phylogenetic tree, alternative splicing and coupling transcriptome. We conducted a systematic study on phylogenetic relationship of pig DNMT1 gene, the DNMT1 gene’s alternative splicing situation in the different immune tissues including nasal mucosa, brain lymph nodes, bone marrow, thymus and spleen, and genome-wide transcriptions caused by DNMT1 gene based on the peripheral blood cell model. Preliminary research results are as follows:1. We used mega3.1 to construct the DNMT1 gene’s phylogenetic tree of multiple species, and revealed the pig DNMT1 gene’s relations of differentiation with other species from an evolutionary perspective. Although the phylogenetic genetic distances of DNMT1 gene between various species were different, more conservation between domesticated agricultural animals was found.2. We used the RT-PCR technology to identify the pig DNMT1 gene’s alternative splicing variants including DNMT1 s, DNMT1o1 and DNMT1o2, and systematically examined the expressions of alternative splicing variants of DNMT1 gene at the m RNA levels in the nasal mucosa, lymph nodes, bone marrow, thymus, spleen with the control group. We found that Dnmt1 s, Dnmt1o1, Dnmt1o2 were expressed in various immune tissues, but there were differences of the abundance and proportion between different immune tissues.3. We used porcine peripheral blood cells as an immune cell model and used the porcine Affymetrix gene expression chip, to establish the analysis framework for coupling transcriptome of DNMT1 gene, including detection of DNMT1 probesets, quality control, identification of differentially expressed transcripts, two-way clustering analysis, and GO annotation.4. We identified a total of 161 differentially expressed transcripts based on the two-tail distribution of probe set Ssc.24344.1.S1_a_at, and we identified a total of 135 differentially expressed transcripts covariated with Ssc.24344.1.S1_at.5. A dimension reduction strategy was preliminarily used to investigate the impact of porcine DNMT1 gene expression variation on the gene expression patterns in the peripheral blood cells. We introduced the non-negative matrix factorization method for dimension reduction of gene expression matrix after a nonspecific filtering analysis, and built 10 new factors. The estimations of correlation coefficients and regression coefficients between DNMT1 gene and factor phenotypes have revealed that there was a coupling between expression variation of DNMT1 gene and gene expression pattern of genome.Based on the immunobiology of porcine DNMT1 gene from multiple views, we got some research meaningful results, which accumulated the porcine DNMT1 gene’s based data, and laid the foundation of pig DNMT1 gene’s immune regulation for further and in-depth study.