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GFP as a marker for transient gene transfer and expression in Mycoplasma hyorhinis

文献类型： 外文期刊

作者： Ishag, Hassan Z. A. 1 ; Liu, Maojun 1 ; Yang, Ruosong 1 ; Xiong, Qiyan 1 ; Feng, Zhixin 1 ; Shao, Guoqing 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Natl Res Ctr Engn & Technol Vet Bioprod, Key Lab Vet Biol Engn & Technol, Inst Vet Med,Minist Agr, Nanjing 210014, Jiangsu, Peoples R China

2.Univ Nyala, Coll Vet Sci, Nyala, Sudan

关键词： Mycoplasma hyorhinis;Plasmid;GFP;Expression

期刊名称：SPRINGERPLUS （ 影响因子：1.13； 五年影响因子：1.29 ）

ISSN： 2193-1801

年卷期： 2016 年 5 卷

页码：

收录情况： SCI

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 similar to 4 x 10(-3) cfu/mu g 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.

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