本研究构建了一个pMD18-O / MHRgfp质粒，该质粒包含p97基因启动子、复制起点、四环素抗性标记和由p97基因启动子控制的GFP基因。将质粒转化到猪鼻支原体中，PCR检测质粒表达情况；将猪鼻支原体置于包含四环素的琼脂中培养，挑选单个克隆检测发现其呈现绿色荧光。该研究表明： GFP可用于跟踪猪鼻支原体的瞬时基因转化和表达，从而简化基因转化和表达的筛选程序。
外文摘要：Mycoplasma hyorhinis (M. hyorhinis) is an opportunistic pathogen of pigs and has been shown to transform cell cultures, which has increased the interest of researchers. The green florescence proteins (GFP) gene of Aquorea victoria, proved to be a vital marker to identify transformed cells in mixed populations. Use of GFP to observe gene transfer and expression in M. hyorhinis (strain HUB-1) has not been described. We have constructed a pMD18-O/MHRgfp plasmid containing the p97 gene promoter, origin of replication, tetracycline resistance marker and GFP gene controlled by the p97 gene promoter. The plasmid transformed into M. hyorhinis with a frequency of ~4 × 10-3 cfu/µg plasmid DNA and could be detected by PCR amplification of the GFP gene from the total DNA of the transformant mycoplasmas. Analysis of a single clone grown on KM2-Agar containing tetracycline, showed a green fluorescence color. Conclusively, this report suggests the usefulness of GFP to monitor transient gene transfer and expression in M. hyorhinis, eventually minimizing screening procedures for gene transfer and expression.
外文关键词：Expression；GFP；Mycoplasma hyorhinis；Plasmid
作者：Ishag HZA, Liu MJ, Yang RS,等。
作者单位：江苏省农业科学院。
期刊名称：SPRINGERPLUS
期刊影响因子：0.982
出版年份：2016
出版刊次：6
点击下载：GFP标记猪鼻支原体基因瞬时转染和表达的研究