Construction of transgenic mice containing human mutant p53 and EB virus latent membrane protein 1

    HE Yingª²Chun, TIAN Daoª²Fa, LU Fangª²Guo, MAO Jiª²Fang

    1Department of Basic Medicine, Hunan College of Traditional Chinese Medicine, Changsha 410007, China, 2Shanghai Nanfang Research Center for Model Organisms, Shanghai 201203, China

    ¡¾Abstract¡¿ AIM: To establish biª²transgenic mice models containing mutation p53 gene and Epsteinª²Barr (EB) virus latent membrane protein 1 gene (LMP1) and to provide reliable tools for studying the reciprocity of mutant p53 with LMP1 in the tumorogenesis process of nasopharyngeal precancerosis. METHODS: Molecular cloning techniques were used to construct the recombinant plasmid pLMP1ª²p53mt containing mutant p53 gene and EB virus LMP1 gene. Linear recombinant plasmid pLMP1ª²p53mt was transfected into mouse androgenesis of germ cells by microinjection and then survival germ cells treated were planted into fallopian tubes of artificial pregnant female mice. Threeª²week filial generation mice were screened by PCR and further identified by southern blot, and the histopathological characteristics of different tissues in 4ª²month transgenic mice were observed by Hª²E staining. RESULTS: Seven hundred and seven mouse zygotes were treated with transgenic vectors DNA by microinjection and then transplanted into 30 artificial pregnant female mice. Twentyª²six of these mice were gravid. The transplantation rate was 86.67%. A total of 179 F0 mice were obtained, of which 9 were carried p53mt gene and LMP1 gene screened by PCR. The positive rate of transgenic mice was 5.03% (9/179). The 9 positive F0 mice carrying foreign genes were further confirmed by southern blot. The rhinal mucosa, nasopharyngeal tissues and oesophagus of one founder had inflammation and its nasopharynx mucosa had focal hyperplasia. CONCLUSION: Transgenic mouse models integrating human mutant p53 gene and LMP1 and with focal hyperplasia of nasopharynx mucosa have been established by microinjection, which provides an effective model in studying the mechanism of nasopharyngeal precancerous lesions. ......