BACKGROUND Although the presences of scrapie associated fibril in the brain tissues is a ultrastructural hallmark for prion diseases, the exact morphological structure of prion during the progression of the disease is still unclear. The host prion protein (PrP) is encoded by PrP gene (PRNP) locating on the chromosome 20 in human and the chromosome 2 in mouse. Recently, a novel correlative light and electron microscopy with Mini Singlet Oxygen Generator (miniSOG) was generated. MiniSOG, a small protein of 106 amino acids, can absorb blue light and emit green fluorescence that is detectable under the fluorescence microscope. MiniSOG can also partially catalyze the polymerization of DAB to form black stained structures in the presence of osmium tetroxide, which is able to be observed under transmission electron microscope. NEW METHODS Two kinds of miniSOG-PrP expressing recombinant plasmids were generated. Correlative photooxidation and transmission electron microscope were used to detect these plasmids. The plasmids were microinjected into fertilized FVB/NJ eggs and Tg mice expressing miniSOG-PrP fusion proteins were selected after successive bred withPRNP KO Tg mice. RESULTS Those two strains of Tg mice, TgSOG23 and Tg231SOG, developed normally and maintained healthy without detectable abnormality after one-year observation. Western blots and immunohistochemical assays with PrP- and miniSOG-specific antibodies confirmed that the chimeric miniSOG-PrP proteins were expressed in the brain tissues of Tg mice. Digital PCR assays proposed that the copy numbers of the inserted external gene in TgSOG23 and Tg231SOG were 2 and 12, respectively. COMPARISON WITH EXISTING METHOD(S) Compared with GFP tag miniSOG is significantly smaller, which makes it easy be operated experimentally and possibly has less influence on the biological function of the labeled protein. Additionally, GFP tag is an ideal marker for immunofluorescent assays, but may not be suitable for ultrastructural assays for prion morphology. CONCLUSION Those Tg mice may supply novel and useful experimental animals for further study on the potential morphological structure formation and deposits of prion in the brain tissues during prion infection.

中文翻译：

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两种表达嵌合MiniSOG-MusPrP的转基因小鼠品系的生成和表征。

背景技术尽管脑组织中瘙痒病相关原纤维的存在是病毒疾病的超微结构特征，但是在疾病发展过程中of病毒的确切形态结构仍不清楚。宿主病毒蛋白（PrP）由位于人类20号染色体和小鼠2号染色体上的PrP基因（PRNP）编码。最近，使用微型单重态氧气发生器（miniSOG）产生了一种新型的相关光电子显微镜。MiniSOG是一种106个氨基酸的小蛋白质，可以吸收蓝光并发出绿色荧光，在荧光显微镜下可以检测到绿色荧光。MiniSOG还可以在四氧化存在下部分催化DAB的聚合反应，形成黑色染色的结构，这可以在透射电子显微镜下观察到。新方法产生了两种表达miniSOG-PrP的重组质粒。用相关的光氧化和透射电子显微镜检测这些质粒。将质粒显微注射到受精的FVB / NJ卵中，并与PRNP KO Tg小鼠连续繁殖后，选择表达miniSOG-PrP融合蛋白的Tg小鼠。结果这两种Tg小鼠TgSOG23和Tg231SOG正常发育并保持健康，经过一年的观察，没有发现异常。用PrP和miniSOG特异性抗体进行的蛋白质印迹和免疫组织化学分析证实，嵌合的miniSOG-PrP蛋白在Tg小鼠的脑组织中表达。数字PCR分析表明，TgSOG23和Tg231SOG中插入的外部基因的拷贝数分别为2和12。与现有方法的比较miniSOG与GFP标记相比要小得多，这使其易于实验操作，并且对标记蛋白的生物学功能影响较小。此外，GFP标签是免疫荧光测定的理想标记，但可能不适用于for病毒形态的超微结构测定。结论那些Tg小鼠可提供新颖有用的实验动物，以进一步研究during病毒感染期间脑组织中potential病毒的潜在形态结构形成和沉积。但可能不适用于病毒形态的超微结构测定。结论那些Tg小鼠可提供新颖有用的实验动物，以进一步研究病毒感染过程中脑组织中病毒的潜在形态结构形成和沉积。但可能不适用于病毒形态的超微结构测定。结论那些Tg小鼠可提供新颖有用的实验动物，以进一步研究病毒感染过程中脑组织中病毒的潜在形态结构形成和沉积。