PrP^c重组蛋白脑内接种金黄地鼠对mRNA表达的影响  

Influence of recombinant bovine PrP^c protein on hamster mRNA expression by injection in brain

作　　者：杨建民[1] 赵德明[1] 李宁[2] 吴长德[1] 宁章勇[1] 郝永新[1] 韩彩霞[1] 

机构地区：[1]中国农业大学国家动物海绵状脑病实验室,北京100094 [2]中国农业大学农业生物技术国家重点实验室,北京100094

出　　处：《中国农业大学学报》2005年第6期27-32,共6页Journal of China Agricultural University

基　　金：国家自然科学基金资助项目(30400325;30571399)

摘　　要：为明确将牛PrPc重组蛋白接种金黄地鼠脑内是否引起异常临床表现、脑组织病理学变化及对其mRNA表达产生影响,为进一步研究PrPc蛋白与异构体PrPsc的结构转换机制提供基础数据,将纯化的牛PrPc重组蛋白磷酸盐缓冲液制剂进行地鼠颅内接种,约3μL/只,对照接种磷酸盐缓冲液(PBS);102 d后取出脑组织:一部分福尔马林固定后进行常规H.E.染色观察,另一部分提取脑组织总RNA,进行实时荧光定量RT-PCR检测。结果表明:处理未见临床异常表现;大脑、小脑、脑干组织病理学检测未见海绵状空泡变性和淀粉样斑;大脑PrPcmRNA表达水平无显著性差异。上述结果表明,用PrPc重组蛋白接种,在检测时间102 d内未引起金黄地鼠行为和脑组织的异常改变。To observe biological specificity of recombinant bovine PrPc protein, such as the pathology to hamster and influence on PrPc gene expression level, a purified prion recombinant structure protein （10μg/uL, 3μL） was injected in the brain of six hamsters, Two hamsters injected with phosphatic salt buffer served as controls. The animals were sacrificed by bleeding through eyeball vein on day 102. Samples consisted of brain, cerebellum, and obex. A part of brain was fixed in formaldehyde, embedded in paraffin wax, sectioned, stained with H.E. and examined under microscope. Another part was used for total RNA extraction. PrPc expression was tested using real-time RT-PCR in both test and control groups. The results indicated that, within 102days, neither abnormal clinical symptoms nor histopathological changes and PrPc gene expression difference were observed between the test and control group. This study has demonstrated that the recombinant protein failed to cause pathological change and PrPc gene expression difference in tested rats. This finding provides a significant information on the prion structural biology based on the recombinant protein.

关 键 词：传染性海绵状脑病 Prion蛋白 实时荧光定量RT-PCR MRNA表达水平 

分 类 号：Q3-3[生物学—遗传学] S865.14[农业科学—野生动物驯养]