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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。
Objective To construct recombinant adenovirus containing transforming growth factor β1 (TGF-β1) antisense RNA. Methods The 630 bp fragment of TGF-β1 cDNA was inserted into the shuttle vector pAdTrack-CMV to construct the recombinant pAdTrack-antiTGFβ1 and the recombinant plasmid pAdEasy- 1 co-transfection BJ5183 bacteria. The positive clones of homologous recombination were screened with selective medium and 293 cells were transfected with homologous recombination adenovirus vector. The recombinant adenovirus was identified by fluorescent microscopy of green fluorescent protein (GFP) and PCR amplification of the target gene virus. Results The recombinant adenovirus containing TGF-β1 antisense RNA was constructed with a titer of 2.4 × 109 PFU / ml. Conclusion The recombinant adenovirus has certain application prospects in the treatment of glomerulonephritis and glomerulosclerosis, and provides a research method for the treatment and research of glomerulosclerosis.