Cloning, Expression Profiles Of The Ovine RPS23, ACAT1 And HPSE Genes And SNPs Association With Hematologic Parameters

This study related to Ribosomal protein S23(RPS23), Acyl-coenzyme A:cholesterolacyl transferase 1(ACAT1) and Heparanase(HPSE) as the research object of immune gene by using the combination of bioinformatics and molecular biology method. We separate the genes, analyse the gene expression and developmental expression, SNP scanning and its associated with hematologic parameters. The results are in the following:We obtained the RPS23 gene whole length Coding sequence of sheep and a part of length of ACAT1 gene and HPSE gene coding sequence, which was 460 bp, 1302 bp and 1708 bp respectively. The RPS23 gene probably functioned as astructural protein in the stress response, signal transduction, immune response and may played an important role in the process of transcription regulation. The ACAT1 probably functioned as a transporter in the signal transduction, stress response, immune response and may played an important role in the process of transcription regulation, which had great effect to thegrowth metabolism of sheep.The HPSE gene probably functioned as a structural protein in the signal transduction, stress response and transcription regulation may played an important role in the process of signal transduction.RPS23, ACAT1 and HPSEgene has a broad expression in 15 tissues of sheep including rumen, reticulum, omasum, abomasum, duodenum, caecum, hypothalamus, hypophysis, heart, liver, spleen, lung, kidney, muscle and fat. RPS23 gene expressed the highest in lung, spleen followed. ACAT1 gene expressed the highest in kidney. HPSE gene expressed the highest in spleen.It did not show obvious regularity of sheep RPS23 m RNA and HPSE m RNA developmental expression in lung and spleen, and the corresponding time point of lung and spleen weight had no significant correlation(P>0.05). The ACAT1 gene expression of 0-84 day sheep raised with the growth of the age in the overall, and its expression increased with the increase of age. Also the weight of the kidney has significantly positive correlation with the time points. The linear regression formula is : Y=0.2311X+0.7813(n=7,R2=0.7828,P=0.015)We use PCR-RFLP and sequencing for SNP scanning and its association with blood index analysis. The results show that there is a c.720A>G BsaⅠrestriction enzyme cutting site of RPS23 gene which polymorphisms is significant correlation with platelet volume distribution width(PDW)(P<0.05); And a g.1060G>A Fnu4HⅠrestriction enzyme cutting site polymorphisms of HPSE gene is significant correlation with acerage red blood cell hemoglobin concentration(MCHC)(P<0.05).