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Transgenic mice with overexpression of mutated human optineurin(E50K) in the retina

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Abstract

In the present work, Site-directed mutagenesis to insert the Glu50Lys amino acid substitution was achieved by PCR using plasmid pBluescript-OPTN. Mutated human OPTN(E50K) gene-driven mouse c-kit promoter was constructed and confirmed by endonuclease digestion and sequence analysis. Transgenic mice were generated via the microinjection method. PCR and DNA dot blot were used to screen the positive transgenic mice. RT-PCR analyzed the RNA level and location of mutated human OPTN(E50K) mRNA expression in transgenic mice. Western blot and immunohistochemistry were used to detect the level and location of mutated human OPTN(E50K) expression in transgenic mice. A transgenic mouse model with overexpression of mutated human OPTN(E50K) in retina was successfully established. The transgene was integrated and transmitted into the chromosome of transgenic mice. Mutated human OPTN(E50K) gene was controlled by c-kit promoter and expressed in the retina in mice. Mutated human OPTN(E50K) in transgenic mice was higher than that of wild type C57BL/6J mice. Our studies had provided a new transgenic model for investigating the molecular properties of mutated human OPTN(E50K).

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Acknowledgments

The authors are grateful to PHD. Hong Cai (Harbin veterinary research institute, CAAS) for his technical help. This project was funded by National Natural Science foundation of China (No. 81070730) and Key Program of Natural Science foundation of Heilongjiang province (ZD201015). China, patent 201110143751.5, 2011.

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Author notes

  1. Qingfeng Meng

    Present address: Department of Ophthalmology, First Affiliated Hospital of Harbin Medical University, Harbin, China

Authors and Affiliations

  1. Department of Ophthalmology, Second Affiliated Hospital of Harbin Medical University, No. 246 Xuefu street, Nangang district, Heilongjiang, Harbin, 150086, China

    Qingfeng Meng, Zheng Xiao, Huiping Yuan, Xinrong Zhou, Binbin Yang, Jingbo Sun, Bo Meng, Xian Sun & Fang Cheng

  2. Division of Livestock Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of the Chinese Academy of Agricultural Sciences, Harbin, 150001, China

    Fei Xue & Yuanmao Zhu

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  1. Qingfeng Meng
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Correspondence to Huiping Yuan.

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Qingfeng Meng and Zheng Xiao contributed equally to this work

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Meng, Q., Xiao, Z., Yuan, H. et al. Transgenic mice with overexpression of mutated human optineurin(E50K) in the retina. Mol Biol Rep 39, 1119–1124 (2012). https://doi.org/10.1007/s11033-011-0840-0

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  • DOI: https://doi.org/10.1007/s11033-011-0840-0

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