【摘要】：本研究克隆了綿羊核糖體蛋白S23基因(Ribosomal protein S23,RPS23)、�；o酶A㑳膽固醇�；D(zhuǎn)移酶1基因(Acyl-coenzyme A:cholesterolacyl transferase 1,ACAT1)和乙酰肝素酶(heparanase,HPSE)基因,研究了其組織表達(dá)模式和0~84d的時序性表達(dá)模式,分析了SNPs與血液指標(biāo)關(guān)聯(lián)性。結(jié)果如下:1、分離得到了綿羊RPS23基因完整編碼區(qū)序列、綿羊ACAT1基因和綿羊HPSE基因的部分編碼區(qū)序列,長度分別為460 bp、1302bp和1708bp;預(yù)測綿羊RPS23基因可能作為一種能作為結(jié)構(gòu)蛋白在信號傳導(dǎo)、脅迫應(yīng)答、免疫應(yīng)答和轉(zhuǎn)錄調(diào)控過程中發(fā)揮重要作用;ACAT1可能作為運(yùn)載體在信號傳導(dǎo)、轉(zhuǎn)錄調(diào)控和免疫應(yīng)答過程中發(fā)揮重要作用;HPSE可能在信號傳導(dǎo)、脅迫應(yīng)答和轉(zhuǎn)錄調(diào)控過程中發(fā)揮重要作用。2、綿羊RPS23、ACAT1和HPSE基因在所研究的15種組織中都有表達(dá),屬廣譜表達(dá)基因,RPS23、ACAT1和HPSE基因分別在肺臟、腎臟和脾臟中表達(dá)量最高。3、綿羊RPS23 mRNA和HPSE mRNA在0~84 d羔羊肺臟和脾臟中的時序性表達(dá)未呈現(xiàn)出明顯的規(guī)律性變化,且與相應(yīng)時間點肺臟和脾臟組織重量無顯著相關(guān)(P0.05)。ACAT1 mRNA在0-84 d羔羊腎臟組織中的時序性表達(dá)總體呈上升趨勢,其表達(dá)量隨日齡的增長而增加,且與相應(yīng)時間點腎臟的重量呈線性顯著的正相關(guān),回歸公式為:Y=0.2311X+0.7813(n=7,R2=0.7828,P=0.015)。4、運(yùn)用PCR-RFLP和測序的方法對綿羊RPS23、ACTA1和HPSE基因進(jìn)行了SNP掃描及其與血液指標(biāo)關(guān)聯(lián)分析。結(jié)果表明,分別在RPS23和HPSE基因中各發(fā)現(xiàn)了1個新的多態(tài)位點,RPS23 c.720AG和HPSE g.1060GA。RPS23c.720AG多態(tài)性與血小板體積分布寬度(PDW)存在顯著相關(guān)(P0.05)。HPSE g.1060GA多態(tài)性與平均紅細(xì)胞血紅蛋白濃度(MCHC)存在顯著相關(guān)(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).
【學(xué)位授予單位】：甘肅農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S826

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