绵羊RPS23、ACAT1和HPSE基因的克隆与组织表达及其SNPs与血液指标关联分析
发布时间：2018-09-03 19:21
【摘要】：本研究克隆了绵羊核糖体蛋白S23基因(Ribosomal protein S23,RPS23)、酰基辅酶A㑳胆固醇酰基转移酶1基因(Acyl-coenzyme A:cholesterolacyl transferase 1,ACAT1)和乙酰肝素酶(heparanase,HPSE)基因,研究了其组织表达模式和0~84d的时序性表达模式,分析了SNPs与血液指标关联性。结果如下:1、分离得到了绵羊RPS23基因完整编码区序列、绵羊ACAT1基因和绵羊HPSE基因的部分编码区序列,长度分别为460 bp、1302bp和1708bp;预测绵羊RPS23基因可能作为一种能作为结构蛋白在信号传导、胁迫应答、免疫应答和转录调控过程中发挥重要作用;ACAT1可能作为运载体在信号传导、转录调控和免疫应答过程中发挥重要作用;HPSE可能在信号传导、胁迫应答和转录调控过程中发挥重要作用。2、绵羊RPS23、ACAT1和HPSE基因在所研究的15种组织中都有表达,属广谱表达基因,RPS23、ACAT1和HPSE基因分别在肺脏、肾脏和脾脏中表达量最高。3、绵羊RPS23 mRNA和HPSE mRNA在0~84 d羔羊肺脏和脾脏中的时序性表达未呈现出明显的规律性变化,且与相应时间点肺脏和脾脏组织重量无显著相关(P0.05)。ACAT1 mRNA在0-84 d羔羊肾脏组织中的时序性表达总体呈上升趋势,其表达量随日龄的增长而增加,且与相应时间点肾脏的重量呈线性显著的正相关,回归公式为:Y=0.2311X+0.7813(n=7,R2=0.7828,P=0.015)。4、运用PCR-RFLP和测序的方法对绵羊RPS23、ACTA1和HPSE基因进行了SNP扫描及其与血液指标关联分析。结果表明,分别在RPS23和HPSE基因中各发现了1个新的多态位点,RPS23 c.720AG和HPSE g.1060GA。RPS23c.720AG多态性与血小板体积分布宽度(PDW)存在显著相关(P0.05)。HPSE g.1060GA多态性与平均红细胞血红蛋白浓度(MCHC)存在显著相关(P0.05)。
[Abstract]:In this study, sheep ribosomal protein S23 gene (Ribosomal protein S23 RPS23, acyl coenzyme A? Cholesterol acyltransferase 1 gene (Acyl-coenzyme A:cholesterolacyl transferase 1 ACA T1) and heparinase (heparanase,HPSE) gene were used to study the expression pattern of cholesterol acyltransferase 1 (Acyl-coenzyme A:cholesterolacyl transferase 1) and heparinase (heparanase,HPSE). The expression patterns of cholesterol acyltransferase 1 (Acyl-coenzyme A:cholesterolacyl transferase 1) and heparinase (heparanase,HPSE) were studied, and the correlation between SNPs and blood index was analyzed. The complete coding region sequence of sheep RPS23 gene, the partial coding region of sheep ACAT1 gene and sheep HPSE gene were obtained as follows: 1. The length is 460 bp,1302bp and 1708 bprespectively. It is predicted that sheep RPS23 gene may play an important role as a structural protein in signal transduction, stress response, immune response and transcriptional regulation. HPSE may play an important role in signal transduction, stress response and transcriptional regulation. Sheep RPS23,ACAT1 and HPSE genes are expressed in 15 kinds of tissues studied. The expression of RPS23 mRNA and HPSE mRNA in lung, kidney and spleen were the highest. The expression of RPS23 mRNA and HPSE mRNA in sheep showed no obvious regular changes in lung and spleen. There was no significant correlation with the weight of lung and spleen tissues at the corresponding time points (P0.05). The temporal expression of ACAT1 mRNA in kidney tissues of lambs at 0-84 days showed a general upward trend, and the expression of ACAT1 mRNA increased with the increase of age. There was a linear and significant positive correlation between the weight of kidney and the weight of kidney at the corresponding time point. The regression formula was: 1: YT 0.2311X 0.7813 (NR27828P0. 015) .4.The RPS23,ACTA1 and HPSE genes of sheep were scanned by SNP and analyzed by PCR-RFLP and sequencing. The results showed that there was a significant correlation between the polymorphism of RPS23 c.720AG and HPSE g.1060GA.RPS23c.720AG in RPS23 and HPSE gene and the (PDW) of platelet volume distribution width (P0.05). The polymorphism of HPSE g.1060GA was significantly correlated with the mean erythrocyte hemoglobin concentration (MCHC) (P0.05).
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826
本文编号：2220931
[Abstract]:In this study, sheep ribosomal protein S23 gene (Ribosomal protein S23 RPS23, acyl coenzyme A? Cholesterol acyltransferase 1 gene (Acyl-coenzyme A:cholesterolacyl transferase 1 ACA T1) and heparinase (heparanase,HPSE) gene were used to study the expression pattern of cholesterol acyltransferase 1 (Acyl-coenzyme A:cholesterolacyl transferase 1) and heparinase (heparanase,HPSE). The expression patterns of cholesterol acyltransferase 1 (Acyl-coenzyme A:cholesterolacyl transferase 1) and heparinase (heparanase,HPSE) were studied, and the correlation between SNPs and blood index was analyzed. The complete coding region sequence of sheep RPS23 gene, the partial coding region of sheep ACAT1 gene and sheep HPSE gene were obtained as follows: 1. The length is 460 bp,1302bp and 1708 bprespectively. It is predicted that sheep RPS23 gene may play an important role as a structural protein in signal transduction, stress response, immune response and transcriptional regulation. HPSE may play an important role in signal transduction, stress response and transcriptional regulation. Sheep RPS23,ACAT1 and HPSE genes are expressed in 15 kinds of tissues studied. The expression of RPS23 mRNA and HPSE mRNA in lung, kidney and spleen were the highest. The expression of RPS23 mRNA and HPSE mRNA in sheep showed no obvious regular changes in lung and spleen. There was no significant correlation with the weight of lung and spleen tissues at the corresponding time points (P0.05). The temporal expression of ACAT1 mRNA in kidney tissues of lambs at 0-84 days showed a general upward trend, and the expression of ACAT1 mRNA increased with the increase of age. There was a linear and significant positive correlation between the weight of kidney and the weight of kidney at the corresponding time point. The regression formula was: 1: YT 0.2311X 0.7813 (NR27828P0. 015) .4.The RPS23,ACTA1 and HPSE genes of sheep were scanned by SNP and analyzed by PCR-RFLP and sequencing. The results showed that there was a significant correlation between the polymorphism of RPS23 c.720AG and HPSE g.1060GA.RPS23c.720AG in RPS23 and HPSE gene and the (PDW) of platelet volume distribution width (P0.05). The polymorphism of HPSE g.1060GA was significantly correlated with the mean erythrocyte hemoglobin concentration (MCHC) (P0.05).
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826
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