Association Of The Differential Expression Of GPR54 With The Transcription Level Related To Genes In Leukocyte Of Wandong Cattle

The variation in GPR54 gene had some association with bovine puberty.In this study,total RNA was extracted from the leukocytes in blood of Wandong cattle(22cattle),and the SNPs of the GPR54 regulatory region were analyzed for the blood samples of cattle to identify the polymorphic fragment genotypes.The expression of GPR54 in different genotypes was analyzed by real-time quantitative PCR.RNA-Seq technique was used to analyze the GPR54 in different genotypes and its correlation with the transcription level and signal pathway.The results showed that:1.Two SNPs were detected in the promoter region of GPR54 gene in Wangdong cattle,and the three genotypes CCTT(A group),CTCT(B group)and TTCC(C group),respectively.2.The expression level of GPR54 gene in different genotype cattle was detected as CCTT> CTTC>TTCC.The expression level of CTCT type,GPR54 gene was between the two homozygous.3.8 samples were detected by RNA-Seq technology,averagely generating24,136,079 raw sequencing reads and then 23,739,830 clean reads.Compared with the reference gene and the reference genome,the average ratio of clean reads were 59.97%and 91.22%,respectively.4.There were 16719 different genes in A group(CCTT)and B group(CTCT)by RNA-Seq.Among them,6778 genes were up-regulated and 9920 genes were down-regulated.By analyzing the results of differential gene screening,we found a large number of genes related to the initiation of primate.For example,the genes of GABA and NPY had inhibitory effects on puberty;TGF and IGF,and GPR54 / Kiss-1 were secreted by glial cell secretion;and GnRH gene,estrogen receptor gene(ER)and sex hormone tuberculosis globulin gene(SHBG)were related to the start of the initial situation.5.Two pathway of enrichment,including the GnRH signaling pathway and the ESTROGEN signaling pathway,were selected by pathway analysis of the differential gene.The difference gene of the signal pathway was analyzed and the CaM gene was significantly down-regulated(P <0.05).