【作者】 包图雅；

【导师】 曹贵方；

【作者基本信息】 内蒙古农业大学 ， 基础兽医学， 2005， 硕士

【摘要】 本文主要研究蒙古绵羊β-防御素-1(sBD-1)。从蒙古绵羊瘤胃黏膜上皮组织中提取总 RNA,采用 RT-PCR 技术扩增出 sBD-1 的 cDNA,将产物测序,得到该 cDNA包含由 192 个碱基组成的开放读码框(ORF),该 ORF 编码 64 个氨基酸残基的前原防御素,该前原防御素含有β-防御素的特征性结构即六个在特定位置上的保守半胱氨酸残基。我们采用 cDNA 末端快速扩增即 RACE 技术,主要包括锚式 PCR RACE 技术和反向嵌套 PCR RACE 技术分别扩增出蒙古绵羊 sBD-1 的 3ˊ末端和 5ˊ末端,经测序得到 sBD-1 的 cDNA 全长序列。 为了检测 sBD-1 mRNA 在蒙古绵羊体内的可能的表达器官,提取蒙古绵羊的整个消化道、生殖器官,心脏等器官内的总 RNA,通过 RT-PCR 检测 mRNA 的表达。结果显示:sBD-1 mRNA 在整个消化道(除直肠)、气管、脾脏、子宫等管状器官内的黏膜层内有表达,而在实质性器官如心脏、肝脏、肺脏、肾脏、淋巴结、膀胱、卵巢、输卵管中无 sBD-1 mRNA 表达。更多还原

【Abstract】 We mainly study Mongolian sheepβ-defensin-1. Total RNA was extracted from the rumen epithelial tissue of Mongolian sheep and the cDNA encoding sBD-1 was amplified by the reverse transcription PCR( RT-PCR ). The results of cDNA sequencing examined that the sBD-1 cDNA contain a open reading frame(ORF) of 192 bases which encoded a 64 amino acid prepro-peptide and the prepro-peptide contained the β–defensin consensus sequence of six invariantly spaced cysteine residues. We adopt the technic of Rapid Amplication of cDNA Ends.The anchored-PCR-RACE strategy was used to amplify 3ˊend of cDNA and backward nested- PCR-RACE strategy was used to amplify 5ˊend of cDNA.Then,they were identified to encode the full cDNA sequence of cBD-1 In order to examine the organs that express possibly sBD-1 mRNA in a Mongolian sheep, We examined the expression of sBD-1 mRNA in different tissues of the Mongolian sheep by RT-PCR. The result indicates that sBD-1 mRNA was detected in the mucosa of throughout the digestive tract except rectum ( tongue, esophagus, rumen, abomasums, reticulum,omasum,duodenum, jejumum, ileum, colon ), trachea, uterus and spleen, but sBD-1 mRNA wasn’t detected in the heart ,liver, lung, kidney, lympho node, bladder,ovarium,oviduct.更多还原