Screening Of Major Weight Genes Of Sika Deer Velvet Antler

The authentic producing areas of Chinese Sika deer velvet antlers in Jilin Province are mainly distributed in the Dongfeng and Shuangyang regions.Velvet antlers have high efficacy and economic value.The growth of velvet antlers are affected by factors such as heredity,breeding conditions,and environment.In order to screen out the major genes related to velvet antlers weight,the differentially expressed genes(DEGs)of Sika deer velvet antlers with different weight were analyzed by high-throughput sequencing.In this experiment,Sika deer in Dongfeng and Shuangyang areas were used as experimental subjects,velvet antlers blood was collected and subjected to high-throughput sequencing.After DEGs were obtained,analyzed by GO and KEGG enrichment and verified via q RT-PCR,and the weight-related genes were cloned and bioinformatics analyzed to understand the gene sequence characteristics and the structure and function of proteins.(1)The total RNA was extracted from velvet antlers blood,and then the quality of the total RNA was detected,from which the qualified samples were screened and grouped according to the velvet antlers weight and place of origin: Dongfeng high-yield group(DF14,DF15,DF37);Dongfeng low-yield group(DF7,DF24,DF32);Shuangyang high-yield group(SY13,SY14,SY15);Shuangyang low-yield group(SY1,SY18,SY28).(2)The samples were analyzed via high-throughput sequencing.The results showed that a total of 561 DEGs were obtained,of which 84 DEGs were obtained in Dongfeng group: 44 genes were up-regulated and 40 genes were down-regulated,Shuangyang group obtained 480 DEGs: 135 genes were up-regulated and 345 genes were down-regulated.GO analysis showed that these genes enriched a total of 927 items,including 337 items for molecular functions,471 items for biological processes,and 119 items for cellular components,through KEGG analysis,DEGs involved a total of 242 pathways: 106 pathways in the dongfeng Group,235 pathways in the shuangyang group.13 DEGs were verified by q RT-PCR,compared with the low-yield group,the trends of BOLA,IGF2,TRPC6,CLEC12 B,CLIC4 and COL12A1 gene were up-regulated,and the trends of SECTM1,CD96 gene were down-regulated in the dongfeng high-yield,the trends of BOLA,ALAS2,CD96,NUDT7 gene were up-regulated,and the trends of WDFY4,VCAN,ADRA2 A gene were down-regulated in the shuangyang high-yield group.The overall trends were consistent with the sequencing data.The relative expression of the ALAS2 gene in the two regions was higher than that of the light velvet antlers.(3)The ALAS2 gene was cloned and analyzed by bioinformatics,it showed that the CDS region of Sika deer ALAS2 gene was 1785 bp in length,which encoded 594 amino acids,the relative molecular weight of ALAS2 protein was 65.90 k Da,the theoretical isoelectric point was 8.28,and the grand average of hydrophilicity was-0.187,and the instability index was computed to be 41.05.ALAS2 protein was an unstable hydrophilicity and had no signal peptides,no transmembrane regions,the result of subcellular localization was in mitochondria.Moreover,it had AAT-I superfamily core sequence.In the secondary structure of ALAS2 protein,the number of alpha helices was the largest,accounting to 43.6%,followed by random coils,accounting to 37.54%.Phylogenetic tree analysis showed that the Sika deer ALAS2 protein was similar to the Cervus elaphus ALAS2 protein,and the Sika deer had the closest evolutionary relationship with the Cervus elaphus.The blood of Sika deer velvet anlter with different weights was analyzed by high-throughput sequencing,the ALAS2 gene might be a major gene that affected the weight of velvet anlter,which was expected to be a key gene for screening velvet anlter weights.