1-2月龄荷斯坦奶牛不同组织中FMDV受体亚基αv、β3、β6mRNA转录水平的相对定量研究  

Relative Quantification of mRNA Transcription of αv,β3,and β6 Subunits Relevant to FMDV Receptors in Different Tissues of Hostein Heifer by Real-time Quantitative RT-PCR

作　　者：独军政[1] 常惠芸[1] 薛霜[1] 高闪电[1] 丛国正[1] 邵军军[1] 林彤[1] 包慧芳[1] 才学鹏[1] 

机构地区：[1]中国农业科学院兰州兽医研究所/家畜疫病病原生物学国家重点实验室/农业部畜禽病毒学重点开放实验室/国家门蹄疫参考实验室,兰州730046

出　　处：《中国农业科学》2010年第3期605-611,共7页Scientia Agricultura Sinica

基　　金：国家自然科学基金项目(30800833);国家科技支撑计划项目(2006BAD06A03);国家重点基础研究发展计划(“973”)项目(2005CB523201)

摘　　要：【目的】建立检测牛口蹄疫病毒(FMDV)受体亚基αv、β3、β6 mRNA相对转录水平的实时定量PCR方法,分析受体αvβ3和αvβ6在不同器官组织中的转录水平。【方法】在对牛FMDV受体基因克隆和测序的基础上,设计实时定量PCR引物,以GAPDH为内参基因,采用△△Ct相对定量PCR方法检测分析FMDV受体亚基αv、β3、β6 mRNA在1—2月龄荷斯坦奶牛体内不同器官组织中的转录表达谱。【结果】αvβ3在荷斯坦奶牛24种组织中均有不同程度的转录表达;αvβ6在甲状腺、硬腭、鼻内皮、喉头、肺、食管、肾、后蹄冠状带等组织表达量较高,在唇、舌皮、软腭、气管、心脏、瘤胃、直肠、前蹄冠状带等组织转录表达量适中,在唾液腺、下颌淋巴结、肝、脾、肌肉等组织未检测到表达;αvβ6在牛体内的表达分布与FMDV组织嗜性基本一致,αvβ6受体可能是决定FMDV组织嗜性的主要功能受体,αvβ3的组织分布似乎与FMDV组织嗜性无关,但不能排除αvβ3在病毒感染过程中的作用。【结论】建立了检测FMDV受体亚基mRNA在不同器官组织中表达水平的相对实时定量PCR方法。[Objective] To establish a method for the detection of mRNA transcription of αv, β3, β6 subunits by real-time quantitative RT-PCR and analyze the transcription profiles of αvβ3 and αvβ6 as FMDV receptors in different tissues of Hostein heifer [ Method ] According to the sequences of αv, β3, and β6 cDNAs, the real-time PCR primers were designed and synthesized. The △△Ct relative quantification method was used to detect the transcription levels of αv, β3, and β6 mRNAs in 24 different tissues of Hostein heifer. [Result] Results of the study showed that the αv and β3 mRNA were transcripted at high or low level in the 24 different tissues. In constrast, the β6 mRNA is only restricted in the partial tissues, and at high level in the thyroid, hard palate, nasal epithelium, laryngeal, lung, esophageal, kidney and at moderate level in the lip skin, soft palate, tracheal, heart, rumen, rectum, coronary band of front leg, and not detected in the salivary gland, neck lymph node, liver, speen and tousle. The distribution pattern is remarkably similar to the tissue tropism of FMDV. The results suggested that the αvβ6 may serve as the major receptor that determines the tissue tropism of FMDV. The tissue distribution pattern of αvβ3 seems to be unrelated to the tissue tropism of FMDV, but the role for αvβ3 as a receptor during FMDV infection can not be ruled out completely. [Conclusion] The △△Ct relative quantitative RT-PCR method was established successfully for the detection of mRNA transcription of αv, β3 and β6 subunits.

关 键 词：FMDV受体 mRNA转录水平 SYBR Green I 荷斯坦奶牛 

分 类 号：S858.23[农业科学—临床兽医学]