【作者】 汤华淳；

【导师】 龙良启；

【作者基本信息】 华中农业大学 ， 基础兽医学， 2008， 硕士

【摘要】 超饲养（Overfeeding）引起鹅肝脏代谢发生改变,使得大量脂肪沉积在肝脏中形成肥肝（fatty liver）。但目前鹅肥肝形成的分子机理仍然不清楚。本研究以朗德鹅和溆浦鹅为试验材料,使用了90对引物进行mRNA差异显示分析,总共分析了大约800条cDNA片段,最后,经过检验,共确定了超饲养后的6个差异表达基因,其中,基因9105的表达水平上调了。在mRNA差异显示分析的基础上,并对基因9105进行了预测,预测结果表明基因9105的cDNA序列含有一个1239bp的开放读码框架（ORF）,与鸡和其它物种的转化生长因子β₂基因（transforming growth factor-beta 2,TGF-β₂）同源。编码一个412个氨基酸的蛋白。蛋白质序列存在2个保守的功能域,即一个引导序列（TGF-β前肽）,一个是功能结构域（TGF-β）。该蛋白质与鸡、非洲爪蛙、短尾猊、人、猪、牛、绵羊、小鼠、大鼠的TGF-β₂蛋白分别具有99%、96%、91%、90%、90%、89%、89%、88%、87%的同源性。相对定量RT-PCR结果表明鹅基因9105在多种组织中表达,其表达量在肌胃中最高,其次是肝脏、脾脏和卵巢,然后是子宫、肺和肌肉,最低的是肾脏和腹脂。更多还原

【Abstract】 The liver metabolize of goose was dramatically changed induced by overfeeding, and causing large fats deposited in liver. At present, the molecular mechanism of fatty liver forming is still remaining un-clear.In this study, mRNA differential display technique was applied to isolate the differentially expressed gene between fatty liver and normal liver of Landes and Xupu goose. Ninety primer pairs were used and approximately eight hundred pieces of cDNA segments were analyzed in this study. Eventually six differentially expressed genes were confirmed by using real-time quantitative PCR technique, among which gene 9105 was confirmed as up regulated in both fatty livers. And besides, gene 9105 was predicted by the bio-informatics technique. The result indicated that cDNA fragments of gene 9105 which was obtained though cDNA cloning contained an open reading frame of 1239 bp which encoded a protein of 412 amino acids, and the sequence of this fragment showed homologous with the mRNA transforming growth factor- beta 2 （TGF-β₂） of chicken and other species, and the protein of 412 amino acids contained two putative conserved domains of TGF-β-propeptide and TGF-β, and has high homology with its homologues-chicken 99%, Xenopus laevis 96%, Monodelphis domestica 91%, human 90%, pig 90%, cattle 89%, Ovis aries 89%, mouse 88%, rat 87%. And by semi-quantitative RT-PCR technique the result showed gene 9105 was expressed in manifold tissues. The expressed level is the highest in muscular stomach, secondly in liver, spleen and ovary, thirdly in uterus, lung and muscle, the lowest in abdominal fat and kidney.更多还原