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中国生物工程杂志

China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (10): 27-31    DOI: 10.13523/j.cb.20151004
    
Modeling of Transgenic Mice Specifically Expressed HPV16-E6 in Skin
TANG Jun-ming1, ZHAO Yan-ping1, LIU Qi1, SHENG Qing-song2, WU Li-ming1, QIAO Guo-hong1
1. Yixing People's Hospital, Clinical laboratory, Yixing 214200, China;
2. WuxiShenRui Biological Products Company, Wuxi 214200, China
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Abstract  

Objective: To construct the eukaryotic vector for E6 gene of papilloma virus (HPV16) specifically expressed in skin and to identify the mRNA or protein of E6 in transgenic mice. Methods: pINV promoter and HPV16-E6 were amplified by polymerase chain reaction (PCR), and the obtained segments were sequentially digested and inserted to pcDNA3.1(-) vector removed CMV promoter, acquiring dpcDNA3.1(-)-pINV-E6 vector. The transgenic mice were prepared by micro-injection, while the RT-PCR, Western blot and immunochemistry were used to detect the expression of E6 in transgenic mice. Results: The dpcDNA3.1(-)-pINV-E6 vector was correctly connected by sequencing. Then two positive mice carrying exogenous gene were produced among 31 experimental mice. The F1 generation was got by transgenic mice mating with the wild type mice. The E6 mRNA or protein was specifically positive in skin of the transgenic mice by RT-PCR, Western blot or immunochemistry. Conclusion: The pINV-E6 transgenic model mice were successfully built with E6 gene specifically expressed in mouse skin. This would lay a solid foundation for the further study of the role of E6 in cancer.



Key wordsPapilloma virus (HPV)      E6 gene      Specific expression      Transgenic mouse     
Received: 03 April 2015      Published: 25 October 2015
ZTFLH:  Q789  
Cite this article:

TANG Jun-ming, ZHAO Yan-ping, LIU Qi, SHENG Qing-song, WU Li-ming, QIAO Guo-hong. Modeling of Transgenic Mice Specifically Expressed HPV16-E6 in Skin. China Biotechnology, 2015, 35(10): 27-31.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20151004     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I10/27

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