本文選題：Protamine　切入點(diǎn)：山羊　出處：《山西農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：Protamine是一類富含精氨酸和半胱氨酸的高堿性蛋白質(zhì)家族。在大多數(shù)動(dòng)物和植物精子形成的后期合成,替代組蛋白與DNA結(jié)合,使精子基因組壓縮,對(duì)精子穩(wěn)定和功能發(fā)揮有特殊作用。本試驗(yàn)以2、3、5、6、9和12月齡晉嵐絨山羊公羊各3只為試驗(yàn)動(dòng)物,通過熒光定量PCR技術(shù)分析了睪丸和精子中Protamine mRNA表達(dá)特性,并利用免疫組化技術(shù)對(duì)睪丸中Protamine蛋白進(jìn)行了定位,并通過光密度值分析對(duì)其蛋白相對(duì)表達(dá)量進(jìn)行定量研究,旨在研究Protamine)基因在山羊睪丸與精子中的表達(dá)規(guī)律,探究其對(duì)精子結(jié)構(gòu)、功能的影響及其機(jī)制。1.山羊Protamine開放閱讀框長(zhǎng)度156 bp,編碼的蛋白質(zhì)含51個(gè)氨基酸,相對(duì)分子質(zhì)量6.8432 kD,等電點(diǎn)12.23,其中含精氨酸和賴氨酸27個(gè),半胱氨酸7個(gè),磷酸化位點(diǎn)2個(gè)(絲氨酸和蘇氨酸),蛋白質(zhì)二級(jí)結(jié)構(gòu)含有p-折疊以及T-轉(zhuǎn)角結(jié)構(gòu)。2.隨著晉嵐絨山羊的生長(zhǎng)發(fā)育,Protamine mRNA表達(dá)量持續(xù)增加,9月齡和12月齡時(shí)Protamine mRNA表達(dá)量顯著高于2、3、5和6月齡時(shí)的表達(dá)量;山羊高活率(≥0.8)精子中Protamine mRNA的表達(dá)量顯著高于低活率(≤0.5)精子中的表達(dá)量。3.山羊Protamine在長(zhǎng)形精子細(xì)胞和精子中表達(dá),睪丸其他生精細(xì)胞及間質(zhì)細(xì)胞中無表達(dá)。蛋白表達(dá)規(guī)律與mRNA表達(dá)量規(guī)律基本一致。本研究揭示了Protamine對(duì)公山羊繁殖機(jī)能和精子功能有重要作用,其明確的作用機(jī)制需進(jìn)一步研究和闡明。
[Abstract]:Protamine is a family of highly alkaline proteins rich in arginine and cysteine. In the late stage of spermatogenesis in most animals and plants, histone binds to DNA and compresses the sperm genome. In this study, the characteristics of Protamine mRNA expression in testis and spermatozoa were analyzed by fluorescence quantitative PCR with 3 male goats of Jinlan cashmere goat at the age of 2 and 3 at the age of 12 months, and 3 male goats of Jinlan cashmere goat at the age of 12 months, respectively, which had a special effect on the stability and function of spermatozoa. The expression of Protamine protein in testis and spermatozoa of goat was studied by using immunohistochemical technique, and the relative expression of the protein was quantitatively studied by means of optical density value (OSD) analysis, in order to study the expression of Protamine gene in goat testis and spermatozoa. To explore the effect of Protamine on sperm structure and function and its mechanism. 1. The length of goat Protamine open reading frame was 156 BP, the encoded protein contained 51 amino acids, the relative molecular weight was 6.8432 KD, the isoelectric point was 12.23, including 27 arginine and lysine, and 7 cysteine. There were two phosphorylation sites (serine and threonine). The protein secondary structure contained pfold and T- corner structure. With the growth and development of Jinlan Cashmere Goats, the expression of Protamine mRNA continued to increase at 9 and 12 months of age. The expression level was significantly higher than that at 5 and 6 months of age. The expression of Protamine mRNA in goat sperm with high motility (鈮，

本文編號(hào)：1621069