【摘要】 目的 构建含转化生长因子β１（ＴＧＦ－β１）反义ＲＮＡ的重组腺病毒重组体。方法 将ＴＧＦ－β１的 ｃＤＮＡ ５’端６３０ ｂｐ片段反向插入穿梭载体ｐＡｄＴｒａｃｋ－ＣＭＶ，构建为 ＴＧＦ－β１反义 ＲＮＡ的重组体（ｐＡｄＴｒａｃｋ－ａｎｔｉＴＧＦβ１），将重组体ｐＡｄＴｒａｃｋ－ａｎｔｉＴＧＦβ１与包装质粒ｐＡｄＥａｓｙ－１共转染ＢＪ５１８３细菌。用选择培养基筛选同源重组的阳性克隆，同源重组的腺病毒载体转染２９３细胞，在荧光显微镜下观察细胞中的绿色荧光蛋白（ＧＦＰ）及ＰＣＲ扩增目的基因等方法鉴定重组的腺病毒。结果 构建了含ＴＧＦ－β１反义 ＲＮＡ的重组腺病毒，其滴度为 ２．４ × １０９ＰＦＵ／ｍｌ。结论 该重组腺病毒在治疗肾小球肾炎及肾小球硬化的研究中有一定的应用前景，为肾小球硬化的治疗及研究提供了一种研究手段。更多还原

【Abstract】 Objective To construct transforming growth factor beta 1 (TGF-β) antisense RNA via recombinant adenovirus. Methods Partial cDNA of TGF-β1 was inserted into adenoviral vector pAdTrack-CMV and then was recombinated with packaging plasmid pAdEasy-1 in BJ5 183 bacteria. The adenovirus was generated in human 293 cells via homologous recomhination. A series of methods such as polymerase chain reaction (PCR) and fluorescence microscopy was employed to identify the generated recombinant adenovirus. Results Recombinant TGF-β1 antisense adenoviruses were constructed and the titer of virus was generally up to 2. 4 × 1 09 phaque forming units per milliliter (PFU /ml). Conclusion The recombinant adenovirus may be useful in the research and treatment for proliferative glomerulonephritis and sclerosing glomerulonephritis.更多还原