Transcriptome Sequencing And Analysis Of Queshan Black Pig And Yorkshire Pig Spleen Tissue

The immunological difference of pigs not only exist in different breeds,also in different developmental stages.Spleen is the largest peripheral immune organ,and also is the most active immune organ.To study the molecular regulation mechanism of porcine immune gene in different developmental stages and different breeds was analyzed on a genome-wide scale,7 d,90 d,180 d of Yorkshire pig and180 d of Queshan black pig were as the research object,transcriptome sequencing,screening immunization database and bioinformatics analysis technology were taken.The study will provide basis for the regulation mechanism of immune gene and for the study of breeding for disease resistance in pig.The results are as follows:1?The results of m RNA sequencing for Yorkshire spleen tissue in three development stages showed that,the number of differentially expressed genes was the most between 180 d and 7 d,and was 1609;it was the least between 180 d and 90 d,124,and the number between 90 d and 7 d was 1490.GO enrichment analysis showed that the differential genes were mainly enrichment in the terms of biological processes,KEGG analysis showed cell proliferation and immune regulation were mainly pathway.Combining with the immune database,the differential genes of 393,455,10,between 90 d and 7 d,between 180 d and 7 d,between 180 d and 90 d,were located in online immunization database,They are not only differential genes in the three groups,but also related to the immune genes,namely VPREB1,HDAC9,CXCL9,MRC2.2?1283 differential genes were obtained from m RNA sequencing of different breeds,368 of 1283 was the immune-related genes.Comparing with Yorkshire,247 were up-regulated and 1036 were down-regulated in Queshan black pig.GO enrichment analysis showed that the immune system process was the most significantly differerent term in biological process,ECM-receptor interaction and Focal adhesion pathway were also the most significant differerent pathway in KEGG pathways,and they were also associated with the immune function.3?The expression level of EGFL6 gene was significantly higher in Queshan black pig than that in Yorkshire(P < 0.01).CDS of EGFL6 was cloned,the bioinformatics analysis and the expression level was studied.The results showed that the length of CDS was 1665 bp,coding 554 amino acids.EGFL6 was hydrophilic secreted protein and had no transmembrane structure,but had 12 phosphorylation sites.The random curl was the main component of secondary structure.The Q-PCR results showed that the expression level of EGFL6 gene in Queshan black pig is significantly higher than in Yorkshire,except for brain,LD,SF.In Yorkshire,the expression level of EGFL6 gene in 7 d was significantly higher than that in 90 d and 180 d,and it was no significant difference between 90 d and 180 d in Yorkshire,except for spleen,small intestine and brain.Above all,the immunological difference exists in Yorkshire and Queshan black pig,and also in different development stages.The selected immune genes which can become the immune marker gene in this study still need a further study.